Methylobacterium compositions for fungal disease control

ABSTRACT

Compositions comprising  Methylobacterium  with anti-fungal activity, methods for controlling plant pathogenic fungi, and methods of making the compositions are provided.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. patent application Ser.16/467,384, filed Jun. 6, 2019, which is the 371 National Phase ofInternational Patent Application PCT/US2017/065081, filed Dec. 7, 2017,which claims the benefit of U.S. Provisional Patent Application No.62/431,927, filed Dec. 9, 2016, the contents of which are incorporatedherein by reference in their entirety.

SEQUENCE LISTING STATEMENT

A sequence listing containing the file named 53907-171478_ST25.txt whichis 32,768 bytes (measured in MS-Windows®) and created on Dec. 1, 2017,contains 60 nucleotide sequences, is provided herewith via the USPTO'sEFS system, and is incorporated herein by reference in its entirety.

BACKGROUND

One-carbon organic compounds such as methane and methanol are foundextensively in nature, and are utilized as carbon sources by bacteriaclassified as methanotrophs and methylotrophs. Methanotrophic bacteriainclude species in the genera Methylobacter, Methylomonas,Methylomicrobium, Methylococcus, Methylosinus, Methylocystis,Methylosphaera, Methylocaldum, and Methylocella (Lidstrom, 2006).Methanotrophs possess the enzyme methane monooxygenase that incorporatesan atom of oxygen from O₂ into methane, forming methanol. Allmethanotrophs are obligate one-carbon utilizers that are unable to usecompounds containing carbon-carbon bonds. Methylotrophs, on the otherhand, can also utilize more complex organic compounds, such as organicacids, higher alcohols, sugars, and the like. Thus, methylotrophicbacteria are facultative methylotrophs. Methylotrophic bacteria includespecies in the genera Methylobacterium, Hyphomicrobium, Methylophilus,Methylobacillus, Methylophaga, Aminobacter, Methylorhabdus, Methylopila,Methylosulfonomonas, Marinosulfonomonas, Paracoccus, Xanthobacter,Ancylobacter (also known as Microcyclus), Thiobacillus,Rhodopseudomonas, Rhodobacter, Acetobacter, Bacillus, Mycobacterium,Arthobacter, and Nocardia (Lidstrom, 2006).

Most methylotrophic bacteria of the genus Methylobacterium arepink-pigmented. They are conventionally referred to as PPFM bacteria,being pink-pigmented facultative methylotrophs. Green (2005, 2006)identified twelve validated species in the genus Methylobacterium,specifically M. aminovorans, M. chloromethanicum, M. dichloromethanicum,M. extorquens, M. fujisawaense, M. mesophilicum, M. organophilum, M.radiotolerans, M. rhodesianum, M. rhodinum, M. thiocyanatum, and M.zatmanii. However, M. nidulans is a nitrogen-fixing Methylobacteriumthat is not a PPFM (Sy et al., 2001). Methylobacterium are ubiquitous innature, being found in soil, dust, fresh water, sediments, and leafsurfaces, as well as in industrial and clinical environments (Green,2006).

Biotic pressures—the combined effects of pathogens, animals, insects,and weeds—result in an annual loss of 20-40% crop productivity.Additional costs of pest control, reduced crop quality, and fundingresearch to combat these pressures, among other factors, make itdifficult to precisely quantify losses due to crop pests and diseases(Savary et al. 2012, Food Sec. DOI: 10.1007/s12571-012-0200-5).Agricultural biologicals present a novel, exciting, and rapidly emergingoption to combat crop pests and diseases in a manner that complementsand preserves current crop germplasm resistance traits, chemical controloptions, and cultural practices (Schlaeppi and Bulgarelli 2015, MPMI28(3): 212-217. DOI: 10.1094/MPMI-10-14-0334-FI).

Fungal diseases are a major cause of yield and quality loss toagronomically important row crops, including corn, soybeans, wheat,cotton, and canola. Among these diseases, species of ascomycete fungi inthe genera Cercospora, Colletotrichum, Fusarium, and Septoria areparticularly widespread and contribute to significant crop losses.Fusarium species alone caused an estimated 36.2 million bushels/yearloss in soybeans from the years 1994-2010 (Wrather et al. 2010). Thesespecies cause a range of foliar, seedling, and stalk maladies, and,unlike rusts or other diseases caused by obligate biotrophic pathogens,there is no complete genetic resistance to these diseases. Further,chemical controls often must be applied at very precise times tosuccessfully stymie disease progress while maximizing profits andprotecting plant health. The narrow thresholds and windows for controlcan result in incomplete or failed crop protection. Microbialinoculants, which can colonize from the seed and persist on plants andin soil for the full duration of the growing season, offer extendedwindows of protection, can compensate for chemical controlinsufficiencies, and extend the benefits of fungicide through resistancemanagement.

SUMMARY

Provided herein are compositions comprising Methylobacterium thatinhibit growth of a plant pathogenic fungus, methods of using thecompositions to control fungal infections of plants, plant parts, andplants derived therefrom, and methods of making the compositions. SuchMethylobacterium that inhibit growth of a plant pathogenic fungus are incertain instances referred to herein as “Methylobacterium that inhibitplant pathogenic fungi” or, in certain contexts, as simply“Methylobacterium”. In certain embodiments, Methylobacterium thatinhibit growth of a plant pathogenic fungus can be distinguished fromother Methylobacterium that do not inhibit plant pathogenic fungi byassaying for the ability of the Methylobacterium to inhibit fungaldisease in a plant or on an isolated plant part.

Provided herein are compositions comprising a mono- or co-culture ofMethylobacterium that inhibit growth of a plant pathogenic fungus and anagriculturally acceptable excipient and/or an agriculturally acceptableadjuvant. In certain embodiments, the Methylobacterium has at least onegene that encodes a 16S RNA having at least 97%, 98%, 99%, 99.5%, or100% sequence identity to SEQ ID NO: 8; or the Methylobacterium isNLS0109 or a derivative thereof In certain embodiments, theMethylobacterium has in its genome one or more polynucleotide markerfragments of at least 50, 60, 100, 120, 180, 200, 240, or 300nucleotides of SEQ ID NOS: 9-11. In certain embodiments, theMethylobacterium has in its genome one or more marker fragmentscomprising a sequence having at least 98%, 99%, or 99.5% sequenceidentity across the entire length of SEQ ID NOS: 9-11. In certainembodiments, the Methylobacterium is not M. radiotolerans or M. oryzae.In certain embodiments, the plant pathogenic fungus is selected from thegroup consisting of a Blumeria sp., a Cercospora sp., a Cochliobolussp., a Colletotrichum sp., a Diplodia sp., an Exserohilum sp., aFusarium sp., a Gaeumanomyces sp., a Macrophomina sp., a Magnaporthesp., a Microdochium sp., a Peronospora sp., a Phakopsora sp., aPhialophora sp., a Phoma sp., a Phymatotrichum sp., a Phytophthora sp.,a Pyrenophora sp., a Pyricularia sp, a Pythium sp., a Rhizoctonia sp., aSclerophthora, a Sclerospora sp., a Sclerotium sp., a Sclerotinia sp., aSeptoria sp., a Stagonospora sp., a Stenocarpella sp. and a Verticilliumsp. In certain embodiments, the Fusarium sp. is selected from the groupconsisting of Fusarium graminearum, Fusarium verticillioides, Fusariumoxysporum, Fusarium virguliforme, and Fusarium solani. In certainembodiments of any of the aforementioned compositions, the compositioncomprises a solid substance wherein a mono-culture or co-culture ofMethylobacterium is adhered thereto. In certain embodiments, the plantpathogenic fungus is a Rhizoctonia sp. or a Sclerotinia sp. In certainembodiments, the Rhizoctonia sp. is Rhizoctonia solani or Rhizoctoniacerealis. In certain embodiments, the Sclerotinia sp. is Sclerotiniasclerotiorum or Sclerotinia homoeocarpa. In certain embodiments, thecomposition comprises a colloid formed by the solid substance wherein amono-culture or co-culture of Methylobacterium is adhered thereto and aliquid. In certain embodiments, the colloid is a gel. In certainembodiments of any of the aforementioned compositions, the compositionis an emulsion. In certain embodiments of any of the aforementionedcompositions, the Methylobacterium is NLS0109, a derivative thereof, ora Methylobacterium related to NLS0109. In certain embodiments of any ofthe aforementioned compositions, the composition further comprises asecond Methylobacterium strain selected from NLS0017, NLS0020, NLS0021,NLS0042, NLS0064, NLS0066, NLS0089, a derivative thereof, or aMethylobacterium sp. related thereto. In certain embodiments,derivatives of Methylobacterium strains NLS0017, NLS0020, NLS0089 andNL0109 or a Methylobacterium sp. related thereto can be identified in bythe presence of one or more marker fragments selected from the groupconsisting of SEQ ID NO: 9-11, 21-24, 37-39, or 49-51, and can bedistinguished from other related Methylobacterium by PCR analysis usingspecific DNA detection assays that include, but are not limited to, DNAprimer and probe combinations provided herein. Use of any of theaforementioned compositions for coating or partially coating a plantpart (e.g., a seed) to inhibit growth of any of the aforementioned plantpathogenic fungi is also provided herein.

In certain embodiments of any of the aforementioned compositions, theMethylobacterium is NLS0109 or a derivative thereof. In certainembodiments, the Methylobacterium is NLS0109, a derivative thereof, or aMethylobacterium related to NLS0109 and the plant pathogenic fungus is aRhizoctonia sp. or a Sclerotinia sp. In certain embodiments, theMethylobacterium is NLS0109, a derivative thereof, or a Methylobacteriumrelated to NLS0109 and the plant pathogenic fungus is Fusariumgraminearum, Cercospora zeae-maydis, or Colletotrichum graminicola. Incertain embodiments, the Methylobacterium is NLS0109, a derivativethereof, or a Methylobacterium related to NLS0109 and the plantpathogenic fungus is Septoria tritici, Stagonospora nodorum, Pythiumspp., Rhizoctonia solani, a Fusarium spp., Magnaportha grisea,Pyrenophora tritici-repentis, Microdochium nivale, Sclerotiniasclerotiorum, Cercospora sojina, Cercospora kikuchii, Fusarium spp.,Rhizoctonia solani, Fusarium virguliforme, Pythium spp., Rhizoctoniasolani, Gibberella zeae, or a Pythium spp. In any of the aforementionedembodiments, the plant pathogenic fungus that is inhibited can be in itsanamorphic form, its teleomorphic form, or in both its anamorphic formand its teleomorphic forms. In any of the aforementioned embodiments,the composition can comprise a fungal inhibitory concentration of themono- or co-culture of Methylobacterium. Use of any of theaforementioned compositions for coating or partially coating a plantpart (e.g., a seed) to inhibit growth of any of the aforementioned plantpathogenic fungi is also provided herein.

In certain embodiments, the Methylobacterium is NLS0109, a derivativethereof, or a Methylobacterium related to NLS0109 and the plantpathogenic fungus is a Rhizoctonia sp. or a Fusarium sp. and the plantis corn or soybean. In certain embodiments of the methods, the Fusariumsp. is one or more species selected from Fusarium graminearum, Fusariumverticillioides, Fusarium oxysporum, Fusarium virguliforme, and Fusariumsolani, and the Rhizoctonia is Rhizoctonia solani. In some embodiments,Methylobacterium sp. control of fungal infections by Fusarium andRhizoctonia is evidenced by improved germination and emergence of seedfollowing application of the Methylobacterium. In certain embodiments,the Methylobacterium is NLS0109, a derivative thereof, or aMethylobacterium related to NLS0109 and the plant pathogenic fungus is aPythium sp. and the plant is corn or soybean. In certain embodiments ofthe methods, the Pythium sp. is one or more species selected fromPythium torulosum, Pythium oopapillum, Pythium sylvaticum and Pythiumlutarium.

In any of the aforementioned embodiments, the composition can furthercomprise an antifungal compound. In certain embodiments, the antifungalcompound can be an azole, a dithiocarbamate, a strobilurin, aphenylamide, a thiazole-carboxamide, a piperidinyl thiazole isoxazolineor a benzimidazole. In certain embodiments, the azole is ipconazole,tebuconazole, triticonazole, metconazole, prothioconazole, triadimenfon,propiconazole and imazalil. In certain embodiments, the compositionfurther comprises a strobilurin fungicide. In certain embodiments, thestrobilurin fungicide is selected from the group consisting ofazoxystrobin, pyraclostrobin, fluoxastrobin, and trifloxystrobin. Incertain embodiments, the strobilurin fungicide is pyraclostrobin. Use ofany of the aforementioned compositions for coating or partially coatinga plant part (e.g., a seed) to inhibit growth of any of theaforementioned plant pathogenic fungi is also provided herein. Incertain embodiments of any of the aforementioned compositions or uses,the combination of Methylobacterium, crop, pathogen, disease, and/orantifungal compounds is as set forth in Table 4.

Also provided are plants or plant parts that are at least partiallycoated with any of the aforementioned compositions comprising a mono- orco-culture of Methylobacterium. In certain embodiments, theMethylobacterium has at least one gene that encodes a 16S RNA having atleast 97%, 98%, 99%, 99.5%, or 100% sequence identity to SEQ ID NO: 8;or the Methylobacterium is NLS0109 or a derivative thereof. In certainembodiments, the Methylobacterium has in its genome one or morepolynucleotide marker fragments of at least 50, 60, 100, 120, 180, 200,240, or 300 nucleotides of SEQ ID NOS: 9-11. In certain embodiments, theMethylobacterium has in its genome one or more marker fragmentscomprising a sequence having at least 98%, 99%, or 99.5% sequenceidentity across the entire length of SEQ ID NOS: 9-11. In certainembodiments, the at least partially coated plant or plant part is acereal plant or cereal plant part. In certain embodiments, the at leastpartially coated cereal plant is selected from the group consisting of arice, wheat, corn, barley, millet, sorghum, oat, and rye plant. Incertain embodiments, the at least partially coated cereal plant part isselected from the group consisting of a rice, wheat, corn, barley,millet, sorghum, oat, and rye plant part. In certain embodiments, the atleast partially coated plant or plant part is a dicot plant part. Incertain embodiments, the dicot plant or plant part is a soybean, peanut,or tomato plant part. In certain embodiments of any of theaforementioned plants or plant parts, the Methylobacterium in thecomposition was obtained from a plant genus, plant species, plantsub-species, or plant cultivar that is distinct from the genus, species,sub-species, or cultivar of the plant or plant part that is coated withthe composition. Also provided are processed plant products thatcomprise a detectable amount of any of the Methylobacterium of any ofthe aforementioned compositions.

Also provided herein are methods of identifying compositions including,but not limited to, soil samples, plant parts, including plant seeds, orprocessed plant products, comprising or coated with Methylobacterium sp.NLS0109, NL0020, NLS0017 or NLS0089 by assaying for the presence ofnucleic acid fragments comprising at least 40, 50, 60, 100, 120, 180,200, 240, or 300 nucleotides of SEQ ID NO: 9-11, 21-24, 37-39, or 49-51in the compositions. In certain embodiments, such methods can comprisesubjecting a sample suspected of containing Methylobacterium sp.NLS0109, NL0020, NLS0017 or NLS0089 to a nucleic acid analysis techniqueand determining that the sample contains one or more nucleic acidcontaining a sequence of at least about 50, 100, 200, or 300 nucleotidesthat is identical to a contiguous sequence in SEQ ID NO: 9-11, 21-24,37-39, or 49-51, wherein the presence of a sequence that is identical toa contiguous sequence in SEQ ID NO: 9-11, 21-24, 37-39, or 49-51 isindicative of the presence of NLS0109 (SEQ ID NO: 9-11), NL0020 (SEQ IDNO: 21-24) NLS0017 (SEQ ID NO: 37-39) or NLS0089 (SEQ ID NO: 49-51) inthe sample. Such nucleic acid analyses include, but are not limited to,techniques based on nucleic acid hybridization, polymerase chainreactions, mass spectroscopy, nanopore based detection, branched DNAanalyses, combinations thereof, and the like. One example of such anucleic acid analysis is a qPCR Locked Nucleic Acid (LNA) based assaydescribed herein. Such analysis can be used to detect Methylobacteriumstrains present at a concentration (CFU/gm of sample) of 10³, 10⁴, 10⁵,10⁶ or more.

Also provided herein are methods of identifying and/or isolatingMethylobacterium sp. that can inhibit growth of a plant pathogenicfungus by assaying for the presence of nucleic acid sequences containedin SEQ ID NO: SEQ ID NO: 9-11, 21-24, 37-39, or 49-51 in theMethylobacterium sp. In certain embodiments, such methods can comprisesubjecting a candidate Methylobacterium sp. to a nucleic acid analysistechnique and determining that the sample contains one or more nucleicacid containing a sequence of at least about 50, 100, 200, or 300nucleotides that is identical to a contiguous sequence in SEQ ID NO:9-11, 21-24, 37-39, or 49-51, wherein the presence of a sequence that isidentical to a contiguous sequence in SEQ ID NO: 9-11, 21-24, 37-39, or49-51 indicates that the candidate Methylobacterium sp. that can inhibitgrowth of a plant pathogenic fungus. Such nucleic acid analyses include,but are not limited to, techniques based on nucleic acid hybridization,polymerase chain reactions, mass spectroscopy, nanopore based detection,branched DNA analyses, combinations thereof, and the like.

In certain embodiments, a Methylobacterium that is NLS0109, a derivativethereof, or a Methylobacterium related to NLS0109, is used in any of theaforementioned compositions, uses thereof, or methods to inhibit growthof a plant pathogenic fungus, where the use and/or method comprisescontrol of any of the pathogens or diseases, in any of the crops,through any of the modes of application set forth in Table 3. In certainembodiments of any of the aforementioned compositions, uses thereof, ormethods, the Methylobacterium is NLS0109, a derivative thereof, or aMethylobacterium related to NLS0109, the plant pathogenic fungus that isinhibited is Fusarium graminearum, Septoria tritici, Stagonosporanodorum, Rhizoctonia solani, a Fusarium spp., Magnaportha grisea,Pyrenophora tritici-repentis, Microdochium nivale, Blumeria graminis f.sp. tritici, or a Pythium sp., and the plant or plant part is a wheatplant or plant part. In certain embodiments of any of the aforementionedcompositions, uses thereof, or methods, the Methylobacterium is NLS0109,a derivative thereof, or a Methylobacterium related to NLS0109, theplant pathogenic fungus that is inhibited is Cercospora zeae-maydis,Colletotrichum graminicola, Fusarium graminearum, Fusariumverticillioides, Gibberella zeae, Rhizoctonia solani, Stenocarpellamaydis, a Pythium sp., Sclerospora graminicola or Sclerophthoramacrospora, and the plant or plant part is a corn plant or corn plantpart. In certain embodiments of any of the aforementioned compositions,uses thereof, or methods, the Methylobacterium is NLS0109, a derivativethereof, or a Methylobacterium related to NLS0109, the plant pathogenicfungus that is inhibited is Sclerotinia sclerotiorum, Cercospora sojina,Cercospora kikuchii, Fusarium spp., Rhizoctonia solani, Septoriaglycines, Fusarium virguliforme, Pythium spp., Peronospora manshurica,or Phytophthora sojae, and the plant or plant part is a soybean plant orsoybean plant part.

In certain embodiments of any of the aforementioned compositions, usesthereof, or methods, the Methylobacterium is NLS0109, a derivativethereof, or a Methylobacterium related thereto and a secondMethylobacterium selected from NLS0020, NLS0017, NLS0089, a derivativethereof, or a Methylobacterium related thereto, is included in thecomposition. In certain embodiments, the combination of NLS0109 with asecond Methylobacterium selected from NLS0020, NLS0017, NLS0089, aderivative thereof, or a Methylobacterium related thereto that is usedin any of the aforementioned compositions, uses thereof, or methodscomprises any of the combinations of Methylobacterium, crops, pathogens,diseases, and/or modes of application set forth in Table 3. For example,in certain embodiments set forth in Table 3, the Methylobacterium isNLS0109, a derivative thereof, or a Methylobacterium related thereto anda second Methylobacterium is NLS0020, a derivative thereof, or aMethylobacterium related thereto, the crop is wheat, the pathogen isFusarium graminearum, Septoria tritici; Stagonospora nodorum, a Pythiumspp., Rhizoctonia solani, a Fusarium spp., Blumeria graminis f. sp.tritici, Magnaporthe grisea, Pyrenophora tritici-repentis, orMicrodochium nival, and application is to a seed in-furrow; foliar; orany combination thereof, where isolates are used alone or in combinationfor any treatment, including NLS0020 applied in-furrow and NLS0109applied foliar. In certain embodiments, the Methylobacterium is NLS0109,a derivative thereof, or a Methylobacterium related thereto and a secondMethylobacterium selected from NLS0020, NLS0017, NLS0089, a derivativethereof, or a Methylobacterium related thereto, the plant pathogenicfungus that is inhibited is Fusarium graminearum, Septoria tritici,Stagonospora nodorum, Rhizoctonia solani, a Fusarium spp., Magnaporthagrisea, Pyrenophora tritici-repentis, Microdochium nivale, Blumeriagraminis f. sp. tritici, or a Pythium sp., and the plant or plant partis a wheat plant or plant part. In certain embodiments, theMethylobacterium is NLS0109, a derivative thereof, or a Methylobacteriumrelated thereto and a second Methylobacterium selected from NLS0020,NLS0021, NLS0017, NLS0089, a derivative thereof, or a Methylobacteriumrelated thereto, the plant pathogenic fungus that is inhibited isCercospora zeae-maydis, Colletotrichum graminicola, Fusariumgraminearum, Fusarium verticillioides, Gibberella zeae, Rhizoctoniasolani, Stenocarpella maydis, a Pythium sp., Sclerospora graminicola orSclerophthora macrospora, and the plant or plant part is a corn plant orcorn plant part. In certain embodiments, the Methylobacterium isNLS0109, a derivative thereof, or a Methylobacterium related thereto anda second Methylobacterium selected from NLS0020, NLS0064, NLS0017,NLS0089, a derivative thereof, or a Methylobacterium related thereto,the plant pathogenic fungus that is inhibited is Sclerotiniasclerotiorum, Cercospora sojina, Cercospora kikuchii, Fusarium spp.,Rhizoctonia solani, Septoria glycines, Fusarium virguliforme, Pythiumspp., Peronospora manshurica, or Phytophthora sojae, and the plant orplant part is a soybean plant or soybean plant part.

In certain embodiments of any of the aforementioned compositions, usesthereof, or methods, the plant or plant part comprises a fungalinhibitory amount of the Methylobacterium. In certain embodiments, afungal inhibitory amount of the Methylobacterium applied to a plant part(e.g., a seed) is about 1.0×10³, 1.0×10⁴, or 1.0×10⁵ to about 1.0×10⁷ or1.0×10⁸ CFUs of PPFM bacteria/plant part (e.g., a seed). In certainembodiments, the Methylobacterium is heterologous to the plant or plantpart. In certain embodiments of any of the aforementioned plant parts,the plant part is a leaf, a stem, a flower, a root, a tuber, or a seed.

Also provided are methods of making any of the aforementionedcompositions containing the Methylobacterium that inhibit growth of aplant pathogenic fungus that comprise combining a Methylobacterium thatinhibit growth of a plant pathogenic fungus with an agriculturallyacceptable excipient and/or with an agriculturally acceptable adjuvant.In certain embodiments, the Methylobacterium has: (i) at least one genethat encodes a 16S RNA having at least 97%, 98%, 99%, 99.5%, or 100%sequence identity to SEQ ID NO: 8; (ii) has in its genome one or morepolynucleotide marker fragments of at least 50, 60, 100, 120, 180, 200,240, or 300 nucleotides of SEQ ID NOS: 9-11; or (iii) has in its genomeone or more marker fragments comprising a sequence having at least 98%,99%, or 99.5% sequence identity across the entire length of SEQ ID NOS:9-11; or the Methylobacterium is NLS0109 or a derivative thereof. Incertain embodiments of the methods, the Methylobacterium sp. is selectedfrom the group consisting of M. aminovorans, M. extorquens, M.fujisawaense, M. mesophilicum, M. radiotolerans, M. rhodesianum, M.nodulans, M. phyllosphaerae, M. thiocyanatum, and M. oryzae. In certainembodiments of the methods, the Methylobacterium is not M. radiotoleransor M. oryzae. In certain embodiments of the methods, theMethylobacterium is NLS0109, a derivative thereof, or a Methylobacteriumrelated to NLS0109. In certain embodiments of any of the aforementionedmethods, the composition further comprises Methylobacterium strainNLS0017, NLS0020, NLS0021, NLS0042, NLS0064, NLS0066, NLS0089, aderivative thereof, or a Methylobacterium sp. related thereto. Incertain embodiments of any of the aforementioned methods, theMethylobacterium is NLS0109, a derivative thereof, or a Methylobacteriumrelated to NLS0109. In certain embodiments, the plant or plant part is asoybean plant or soybean plant part. In certain embodiments, the plantor plant part is selected from the group consisting of a rice, wheat,corn, barley, millet, sorghum, oat, and rye plant or plant part. Incertain embodiments of the methods, the plant pathogenic fungus isselected from the group consisting of a Blumeria sp., a Cercospora sp.,a Cochliobolus sp., a Colletotrichum sp., a Diplodia sp., an Exserohilumsp., a Fusarium sp., a Gaeumanomyces sp., a Macrophomina sp., aMagnaporthe sp., a Microdochium sp., a Peronospora sp., a Phakopsorasp., a Phialophora sp., a Phoma sp., a Phymatotrichum sp., aPhytophthora sp., a Pyrenophora sp., a Pyricularia sp, a Pythium sp., aRhizoctonia sp., a Sclerophthora, a Sclerospora sp., a Sclerotium sp., aSclerotinia sp., a Septoria sp., a Stagonospora sp., a Stenocarpella sp.and a Verticillium sp.

In certain embodiments of any of the aforementioned compositions, usesthereof, or methods, the mono- or co-culture of Methylobacterium isadhered to a solid substance. In certain embodiments of the methods, theMethylobacterium that is adhered to the solid substance is combined witha liquid to form a composition that is a colloid. In certain embodimentsof the methods, the colloid is a gel. In certain embodiments of themethods, the mono- or co-culture of Methylobacterium adhered to thesolid substance is provided by culturing the Methylobacterium in thepresence of the solid substance. In certain embodiments of the methods,the composition comprises an emulsion. In certain embodiments of themethods, the Methylobacterium is provided by culturing theMethylobacterium in an emulsion. In any of the aforementionedembodiments, the plant pathogenic fungus that is inhibited can be in itsanamorphic form, its teleomorphic form, or in both its anamorphic andteleomorphic forms. In any of the aforementioned embodiments, thecomposition can further comprise an antifungal compound selected fromthe group consisting of an azole, dithiocarbamate, strobilurin, andbenzimidazole. In certain embodiments, the azole is ipconazole.

Also provided are methods for controlling a plant pathogenic fungus thatcomprise applying any of the aforementioned compositions as a firstcomposition that contains one or more Methylobacterium that inhibitgrowth of a plant pathogenic fungus to a plant or a plant part, to soilwhere a plant is grown, to soil where a plant part such as a seed isdeposited, or any combination thereof, in an amount that provides forinhibition of infection by the plant pathogenic fungus in the plant,plant part, or a plant obtained therefrom relative to infection of acontrol plant, plant part, or plant obtained therefrom that had notreceived an application of the composition or been grown in soil treatedwith the composition. In certain embodiments, the Methylobacterium: (i)has at least one gene that encodes a 16S RNA having at least 97%, 98%,99%, 99.5%, or 100% sequence identity to SEQ ID NO: 8; (ii) has in itsgenome one or more polynucleotide marker fragments of at least 50, 60,100, 120, 180, 200, 240, or 300 nucleotides of SEQ ID NOS: 9-11; or(iii) has in its genome one or more marker fragments comprising asequence having at least 98%, 99%, or 99.5% sequence identity across theentire length of SEQ ID NOS: 9-11; or the Methylobacterium is NLS0109, aderivative thereof, or is a Methylobacterium related to NLS0109. Incertain embodiments, the Methylobacterium contains in its genome one ormore marker fragments having a sequence of SEQ ID NOS: 9-11.

In certain embodiments of any of the aforementioned methods or uses, theapplication of the composition provides for at least 40%, 50%, 75%, atleast 85%, or at least 95% inhibition of a plant pathogenic fungalinfection in the plant, plant part, or a plant derived therefromrelative to infection of the control plant, plant part, or plantobtained therefrom. In certain embodiments of the methods, the plantpart is selected from the group consisting of a leaf, a stem, a flower,a root, a tuber, and a seed. In certain embodiments of the methods, themethod further comprises the step of harvesting at least one plant partselected from the group consisting of a leaf, a stem, a flower, a root,a tuber, or a seed from the plant or plant part. In certain embodimentsof the methods, the mycotoxin levels in the plant part are reduced by atleast 50%, at least 75%, at least 85%, or at least 95% relative to aplant part obtained from the control plant, plant part, or plantobtained therefrom. In certain embodiments of the aforementionedmethods, the method further comprises obtaining a processed food or feedcomposition from the plant or plant part. In certain embodiments of theaforementioned methods, the mycotoxin levels in the processed food orfeed composition are reduced by at least 50%, at least 75%, at least85%, or at least 95% relative to a processed food or feed compositionobtained from the control plant, plant part, or plant obtainedtherefrom. In certain embodiments, a fungal inhibitory amount of theMethylobacterium is applied to the plant part. In certain embodiments,the fungal inhibitory amount of the Methylobacterium applied to a plantpart (e.g., a seed) is about 1.0×10³, 1.0×10⁴, or 1.0×10⁵ to about1.0×10⁷, 1.0×10⁸, 1.0×10⁹, or 1.0×10¹⁰ CFUs of Methylobacterium/plantpart (e.g., a seed). In certain embodiments, the Methylobacterium isheterologous to the plant or plant part. In certain embodiments of anyof the aforementioned methods, the plant part is a leaf, a stem, aflower, a root, a tuber, or a seed. In certain embodiments of themethods, the Methylobacterium is NLS0109, a derivative thereof, or aMethylobacterium related to NLS0109. In certain embodiments of any ofthe aforementioned methods, the composition further comprisesMethylobacterium strain NLS0017, NLS0020, NLS0021, NLS0042, NLS0064,NLS0066, NLS0089, a derivative thereof, or a Methylobacterium sp.related thereto. In certain embodiments, a derivative of NLS0020,NLS0017, or NLS0089, or a Methylobacterium sp. related to NLS0020,NLS0017, or NLS0089, can be identified by the presence of one or moremarker fragments having a sequence of SEQ ID NO:21-24, 37-39, or 49-51.In certain embodiments of any of the aforementioned methods, theMethylobacterium is NLS0109 or a derivative thereof. In certainembodiments, the plant or plant part is a soybean plant or soybean plantpart, or a peanut plant or peanut plant part. In certain embodiments,the plant or plant part is selected from the group consisting of a rice,wheat, corn, barley, millet, sorghum, oat, and rye plant or plant part.

In certain embodiments, the methods can further comprise applying asecond composition to soil where the plant is grown, wherein the secondcomposition comprises Methylobacterium strain NLS0017, NLS0020, NLS0021,NLS0042, NLS0064, NLS0066, NLS0089, a derivative thereof, or aMethylobacterium sp. related thereto. In certain embodiments, the secondcomposition is applied to the soil in furrow with the plant part or witha seed from which the plant is grown. In certain embodiments, the firstcomposition is applied to foliage of the plant after application of thesecond composition to the soil. In certain embodiments, the firstcomposition comprises NLS0109, a derivative thereof, or aMethylobacterium sp. related thereto; and the second compositioncomprises Methylobacterium strain NLS0020, NLS0021, NLS0017, aderivative thereof, or a Methylobacterium sp. related thereto. Incertain embodiments, the first composition comprises NLS0109, aderivative thereof, or a Methylobacterium sp. related thereto and astrobilurin fungicide; and the second composition comprisesMethylobacterium strain NLS0020, NLS0021, NLS0017, a derivative thereof,or a Methylobacterium sp. related thereto.

In certain embodiments, the methods can further comprise applying afungicide to a plant, plant part or soil. In such embodiments, thefungicide can be applied simultaneously with one or more of theMethylobacterium sp. or may be applied as a separate composition to aplant or plant part. In certain embodiments, the fungicide is astrobilurin, a phenylamide, a thiazole-carboxamide, or a piperidinylthiazole isoxazoline fungicide. In certain embodiments, the strobilurinfungicide is selected from the group consisting of azoxystrobin,pyraclostrobin, fluoxystrobin, and trifloxystrobin. In certainembodiments, the strobilurin fungicide is pyraclostrobin. In certainembodiments, the fungicide is metalaxyl, mefenoxam, ethaboxam oroxathiapiprolin. In certain embodiments of any of the aforementionedmethods, the combination of Methylobacterium, crop, pathogen, disease,and/or antifungal compounds is as set forth in Table 4. In certainembodiments, the compositions provided herein or the plants or plantparts treated with the compositions can further comprise one or more ofthe aforementioned fungicides.

In certain embodiments, the Methylobacterium is NLS0109, a derivativethereof, or a Methylobacterium related to NLS0109 and the plantpathogenic fungus is a Fusarium sp., a Rhizoctonia sp., a Colletotrichumsp., a Cercospora sp., a Septoria sp., a Pythium sp., a Puccinia sp. ora Sclerotinia sp. In certain embodiments, the Methylobacterium isNLS0109, a derivative thereof, or a Methylobacterium related to NLS0109,the plant pathogenic fungus that is inhibited is Fusarium graminearum,Septoria tritici, Stagonospora nodorum, Rhizoctonia solani, a Fusariumspp., Magnaportha grisea, Pyrenophora tritici-repentis, Microdochiumnivale, Blumeria graminis f. sp. tritici, or a Pythium sp., and theplant or plant part is a wheat plant or plant part. In certainembodiments, the Methylobacterium is NLS0109, a derivative thereof, or aMethylobacterium related to NLS0109, the plant pathogenic fungus that isinhibited is Cercospora zeae-maydis, Colletotrichum graminicola,Fusarium graminearum, Fusarium verticillioides, Gibberella zeae,Rhizoctonia solani, Stenocarpella maydis, a Pythium sp., Sclerosporagraminicola or Sclerophthora macrospora, and the plant or plant part isa corn plant or corn plant part. In certain embodiments, theMethylobacterium is NLS0109, a derivative thereof, or a Methylobacteriumrelated to NLS0109, the plant pathogenic fungus that is inhibited isSclerotinia sclerotiorum, Cercospora sojina, Cercospora kikuchii,Fusarium spp., Rhizoctonia solani, Septoria glycines, Fusariumvirguliforme, Pythium spp., Peronospora manshurica, or Phytophthorasojae, and the plant or plant part is a soybean plant or soybean plantpart.

Also provided are isolated Methylobacterium that inhibit growth of aplant pathogenic fungus. In certain embodiments, the Methylobacteriumsp. that inhibits growth of a plant pathogenic fungus has a 16S nucleicacid sequence having at least 97%, 98%, 99%, 99.5%, or 100% sequenceidentity to SEQ ID NO: 8; or the Methylobacterium is NLS0109 or aderivative thereof. In certain embodiments, the isolatedMethylobacterium has in its genome one or more polynucleotide markerfragments of at least 50, 60, 100, 120, 180, 200, 240, or 300nucleotides of SEQ ID NOS: 9-11. In certain embodiments, the isolatedMethylobacterium has in its genome one or more marker fragmentscomprising a sequence having at least 98%, 99%, or 99.5% sequenceidentity across the entire length of SEQ ID NOS: 9-11. In certainembodiments, the plant pathogenic fungus is selected from the groupconsisting of a Blumeria sp., a Cercospora sp., a Cochliobolus sp., aColletotrichum sp., a Diplodia sp., an Exserohilum sp., a Fusarium sp.,a Gaeumanomyces sp., a Macrophomina sp., a Magnaporthe sp., aMicrodochium sp., a Peronospora sp., a Phakopsora sp., a Phialophorasp., a Phoma sp., a Phymatotrichum sp., a Phytophthora sp., aPyrenophora sp., a Pyricularia sp, a Pythium sp., a Rhizoctonia sp., aStagonospora sp., a Sclerotium sp., a Sclerotinia sp., a Septoria sp., aStagonospora sp., a Stenocarpella sp. and a Verticillium sp.

In any of the aforementioned embodiments, the plant pathogenic fungithat is inhibited can be in its anamorphic form, its teleomorphic form,or in both its anamorphic and teleomorphic forms. The use of any of theaforementioned isolated Methylobacterium to inhibit growth of any one ofthe aforementioned plant pathogenic fungi is also provided herein

BRIEF DESCRIPTION OF THE DRAWINGS

The accompanying drawings, which are incorporated in and form a part ofthe specification, illustrate certain embodiments of the presentdisclosure. In the drawings:

FIG. 1 is a bar chart showing suppression of soybean white mold wiltsymptom severity by NLS0109. White bars represent the mean formock-inoculated controls; gray bars represent the mean for allinoculated plants within each treatment group. Means are calculated fromthree independent experiments with eight replicated pots per experiment(n=24). Errors bars represent±one SEM. Severity was rated on a 0-5scale, with 0=no symptom development and 5=dead plant.

FIG. 2 is a bar chart showing suppression of soybean white mold lesionlength development by NLS0109. White bars represent the mean formock-inoculated controls; gray bars represent the mean for allinoculated plants within each treatment group. Means are calculated fromthree independent experiments with eight replicated pots per experiment(n=24). Errors bars represent±one SEM. Lesion length was measured to thenearest millimeter using a ruler held against the infected plant.

FIG. 3 is a bar chart showing the additive effect on reduction of lategray leaf spot disease severity in corn of NLS0020 in-furrow applicationfollowed by NL0109 foliar application. Applications were made during the2015 field season at a trial site in Bethel, Mo. NLS0020 was appliedin-furrow at a rate of 1,250 mL/A and NLS0109 was applied at a foliarrate of 5,000 mL/A.

FIG. 4 is a line graph showing the progress of gray leaf spot diseaseseverity over time in corn plants treated with PPFM strains, variouscommercial fungicides, and combinations thereof. Groups of ten plantswere treated with various PPFM and fungicide treatments representingstandard disease management practices or alternate PPFM treatmentregimes alone or in combination with conventional chemistries. Diseaseincidence and severity data was collected at 14-, 21-, and 28-dayspost-inoculation and used to calculate disease index values[(incidence*severity)/100].

DESCRIPTION Definitions

As used herein, the phrases “adhered thereto” and “adherent” refer toMethylobacterium that are associated with a solid substance by growing,or having been grown, on a solid substance.

As used herein, the phrase “agriculturally acceptable adjuvant” refersto a substance that enhances the performance of an active agent in acomposition comprising a mono-culture or co-culture of Methylobacteriumfor treatment of plants and/or plant parts.

As used herein, the phrase “agriculturally acceptable excipient” refersto an essentially inert substance that can be used as a diluent and/orcarrier for an active agent in a composition for treatment of plantsand/or plant parts. In certain compositions, an active agent cancomprise a mono-culture or co-culture of Methylobacterium.

As used herein, the phrase “derivatives thereof”, when used in thecontext of a Methylobacterium strain, or isolate refers to any strain orisolate that is obtained from a given Methylobacterium strain.Derivatives of a Methylobacterium strain include, but are not limitedto, variants of the strain obtained by selection, variants of the strainselected by mutagenesis and selection, variants of the strains resultingfrom unselected mutations resulting from growth in culture over one ormore generations, and genetically transformed strains obtained from aMethylobacterium strain.

As used herein, the term “Methylobacterium” refers to bacteria that arefacultative methylotrophs of the genus Methylobacterium. The termMethylobacterium, as used herein, thus does not encompass includesspecies in the genera Methylobacter, Methylomonas, Methylomicrobium,Methylococcus, Methylosinus, Methylocystis, Methylosphaera,Methylocaldum, and Methylocella, which are obligate methanotrophs.

As used herein, the phrase “co-culture of Methylobacterium” refers to aMethylobacterium culture comprising at least two strains ofMethylobacterium or at least two species of Methylobacterium.

As used herein, the term “cultivar” refers to any plant known only incultivation and includes asexually propagated plants, sexuallypropagated plants, inbred lines, and hybrids.

As used herein, the phrase “contaminating microorganism” refers tomicroorganisms in a culture, fermentation broth, fermentation brothproduct, or composition that were not identified prior to introductioninto the culture, fermentation broth, fermentation broth product, orcomposition.

As used herein, the term “emulsion” refers to a colloidal mixture of twoimmiscible liquids wherein one liquid is the continuous phase and theother liquid is the dispersed phase. In certain embodiments, thecontinuous phase is an aqueous liquid and the dispersed phase is liquidthat is not miscible, or partially miscible, in the aqueous liquid.

As used herein, the phrase “essentially free of contaminatingmicroorganisms” refers to a culture, fermentation broth, fermentationproduct, or composition where at least about 95% of the microorganismspresent by amount or type in the culture, fermentation broth,fermentation product, or composition are the desired Methylobacterium orother desired microorganisms of pre-determined identity.

As used herein, the phrase “a fungal inhibitory concentration of themono- or co-culture of Methylobacterium” is a concentration thatprovides for at least a 40%, 50%, 75%, at least 85%, or at least 95%inhibition of a plant pathogenic fungal infection in a plant, plantpart, or a plant derived therefrom relative to infection of the controlplant or plant part.

As used herein, the term “heterologous”, when used in the context ofMethylobacterium that at least partially coats a plant or plant part,refers to a Methylobacterium that is not naturally associated with aplant or plant part of the same species as the plant or plant part thatis at least partially coated with the Methylobacterium. In certainembodiments, the heterologous Methylobacterium that is used to at leastpartially coat a plant or plant part of a first plant species is aMethylobacterium that was isolated, or can be isolated, from a secondand distinct plant species.

As used herein, the phrase “inanimate solid substance” refers to asubstance which is insoluble or partially soluble in water or aqueoussolutions and which is either non-living or which is not a part of astill-living organism from which it was derived.

As used herein, the phrase “a Methylobacterium related to NLS0109” canrefer to a Methylobacterium that: (i) has at least one gene that encodesa 16S RNA having at least 97%, 98%, 99%, 99.5%, or 100% sequenceidentity to SEQ ID NO: 8; (ii) has in its genome one or morepolynucleotide marker fragments of at least 50, 60, 100, 120, 180, 200,240, or 300 nucleotides of SEQ ID NOS: 9-11; or (iii) has in its genomeone or more marker fragments comprising a sequence having at least 98%,99%, or 99.5% sequence identity across the entire length of SEQ ID NOS:9-11.

As used herein, the phrase “mono-culture of Methylobacterium” refers toa Methylobacterium culture consisting of a single strain ofMethylobacterium.

As used herein, a “pesticide” refers to an agent that is insecticidal,fungicidal, nematocidal, bacteriocidal, or any combination thereof.

As used herein, the phrase “bacteriostatic agent” refers to agents thatinhibit growth of bacteria but do not kill the bacteria.

As used herein, the phrase “pesticide does not substantially inhibitgrowth of the Methylobacterium” refers to any pesticide that whenprovided in a composition comprising a fermentation product comprising asolid substance wherein a mono-culture or co-culture of Methylobacteriumis adhered thereto, results in no more than a 50% inhibition ofMethylobacterium growth when the composition is applied to a plant orplant part in comparison to a composition lacking the pesticide. Incertain embodiments, the pesticide results in no more than a 40%, 20%,10%, 5%, or 1% inhibition of Methylobacterium growth when thecomposition is applied to a plant or plant part in comparison to acomposition lacking the pesticide.

As used herein, the term “PPFM bacteria” refers without limitation tobacterial species in the genus Methylobacterium other than M. nodulans.

As used herein, the phrase “solid substance” refers to a substance whichis insoluble or partially soluble in water or aqueous solutions.

As used herein, the phrase “solid phase that can be suspended therein”refers to a solid substance that can be distributed throughout a liquidby agitation.

As used herein, the term “non-regenerable” refers to either a plant partor processed plant product that cannot be regenerated into a wholeplant.

As used herein, the phrase “substantially all of the solid phase issuspended in the liquid phase” refers to media wherein at least 95%,98%, or 99% of solid substance(s) comprising the solid phase aredistributed throughout the liquid by agitation.

As used herein, the phrase “substantially all of the solid phase is notsuspended in the liquid phase” refers to media where less than 5%, 2%,or 1% of the solid is in a particulate form that is distributedthroughout the media by agitation.

To the extent to which any of the preceding definitions is inconsistentwith definitions provided in any patent or non-patent referenceincorporated herein by reference, any patent or non-patent referencecited herein, or in any patent or non-patent reference found elsewhere,it is understood that the preceding definition will be used herein.

Methylobacterium That Inhibit Plant Pathogenic Fungi, CompositionsComprising Methylobacterium That Inhibit Plant Pathogenic Fungi, Methodsof Their Use, and Methods of Making

Various Methylobacterium that inhibit plant pathogenic fungi,compositions comprising these Methylobacterium, methods of using thecompositions to inhibit plant pathogenic fungi, and methods of makingthe compositions are provided herein. As used herein, inhibition of thegrowth of a plant pathogenic fungus includes any measurable decrease infungal growth, where fungal growth includes but is not limited to anymeasurable decrease in the numbers and/or extent of fungal cells,spores, conidia, or mycelia. As used herein, inhibition of infection bya plant pathogenic fungus and/or inhibition of the growth of a plantpathogenic fungus are also understood to include any measurable decreasein the adverse effects caused by fungal growth in a plant. Adverseeffects of fungal growth in a plant include, but are not limited to, anytype of plant tissue damage or necrosis, any type of plant yieldreduction, any reduction in the value of the crop plant product, and/orproduction of undesirable fungal metabolites or fungal growthby-products including, but not limited to, mycotoxins. Plant pathogenicfungi that are inhibited by the compositions and Methylobacteriumprovided herein can be in their anamorphic form, their teleomorphicform, or in both their anamorphic and teleomorphic forms.

Methylobacterium and compositions comprising the same that inhibitgrowth of a plant pathogenic fungus are provided herein. In certainembodiments, the Methylobacterium or composition provides for at leastabout 25%, at least about 40%, at least about 50%, or at least about 75%inhibition of plant pathogenic fungal growth in comparison to a controltreatment upon exposure to a plant pathogenic fungus. In certainembodiments, the plant pathogenic fungus that is inhibited is selectedfrom the group consisting of a Blumeria sp., a Cercospora sp., aCochliobolus sp., a Colletotrichum sp., a Diplodia sp., an Exserohilumsp., a Fusarium sp., a Gaeumanomyces sp., a Macrophomina sp., aMagnaporthe sp., a Microdochium sp., a Peronospora sp., a Phakopsorasp., a Phialophora sp., a Phoma sp., a Phymatotrichum sp., aPhytophthora sp., a Pyrenophora sp., a Pyricularia sp, a Pythium sp., aRhizoctonia sp., a Sclerophthora sp., a Sclerospora sp., a Sclerotiumsp., a Sclerotinia sp., a Septoria sp., a Stagonospora sp., aStenocarpella sp. and a Verticillium sp.

In certain embodiments, the plant pathogenic fungus that is inhibited isa Fusarium sp. In certain embodiments, the Fusarium sp. that isinhibited is selected from the group consisting of Fusarium graminearum,Fusarium verticillioides, Fusarium oxysporum, Fusarium virguliforme, andFusarium solani. In certain embodiments, the isolated Methylobacteriumis NLS0109, a derivative thereof, or a Methylobacterium related toNLS0109. In certain embodiments, Methylobacterium related to NLS0109include, but are not limited to, (i)Methylobacterium sp. that inhibitgrowth of a plant pathogenic fungus and that have a gene encoding a 16SRNA sequence that has at least 97%, 98%, 99%, 99.5%, or 100% sequenceidentity to SEQ ID NO:8; (ii) a Methylobacterium has in its genome oneor more polynucleotide marker fragments of at least 50, 60, 100, 120,180, 200, 240, or 300 nucleotides of SEQ ID NOS: 9-11; or (iii) is aMethylobacterium that has in its genome one or more marker fragmentscomprising a sequence having at least 98%, 99%, or 99.5% sequenceidentity across the entire length of SEQ ID NOS: 9-11. In certainembodiments, the composition further comprises Methylobacterium strainNLS0017, NLS0020, NLS0021, NLS0042, NLS0064, NLS0066, NLS0089, aderivative thereof, or a Methylobacterium sp. related thereto. Plantpathogenic fungi that are inhibited by the compositions andMethylobacterium provided herein can be in their anamorphic form, theirteleomorphic form, or in both their anamorphic and teleomorphic forms.

Also provided are compositions that comprise Methylobacterium thatinhibit growth of a plant pathogenic fungus. In certain embodiments, thecompositions further comprise an agriculturally acceptable excipientand/or an agriculturally acceptable adjuvant. In certain embodiments,the composition provides for at least about 25%, about 50%, or about 75%inhibition of plant pathogenic fungal growth in comparison to a controltreatment upon exposure to a plant pathogenic fungus. In certainembodiments, the plant pathogenic fungus that is inhibited is selectedfrom the group consisting of a Blumeria sp., a Cercospora sp., aCochliobolus sp., a Colletotrichum sp., a Diplodia sp., an Exserohilumsp., a Fusarium sp., a Gaeumanomyces sp., a Macrophomina sp., aMagnaporthe sp., a Microdochium sp., a Peronospora sp., a Phakopsorasp., a Phialophora sp., a Phoma sp., a Phymatotrichum sp., aPhytophthora sp., a Pyrenophora sp., a Pyricularia sp, a Pythium sp., aRhizoctonia sp., a Sclerophthora, a Sclerospora sp., a Sclerotium sp., aSclerotinia sp., a Septoria sp., a Stagonospora sp., a Stenocarpella sp.and a Verticillium sp.

In certain embodiments, the plant pathogenic fungus that is inhibited isa Fusarium sp. In certain embodiments, the Fusarium sp., which isinhibited is selected from the group consisting of Fusarium graminearum,Fusarium verticillioides, Fusarium oxysporum, Fusarium virguliforme, andFusarium solani. In certain embodiments of any of the aforementionedcompositions, the composition comprises a solid substance wherein amono-culture or co-culture of Methylobacterium is adhered thereto. Incertain embodiments where the Methylobacterium is adhered to a solidsubstance, the composition comprises a colloid formed by the solidsubstance wherein a mono-culture or co-culture of Methylobacterium isadhered thereto and a liquid. In certain embodiments, the colloid is agel. In certain embodiments of certain aforementioned compositions,composition is an emulsion that does not contain a solid substance. Incertain embodiments of any of the aforementioned compositions, theMethylobacterium sp. that inhibits growth of a plant pathogenic fungushas a gene encoding a 16S RNA sequence that has at least 97%, 98%, 99%,99.5%, or 100% sequence identity to SEQ ID NO:8. In certain embodimentsof any of the aforementioned compositions, the Methylobacterium isNLS0109, a derivative thereof, or a Methylobacterium related to NLS0109.In certain embodiments of any of the aforementioned compositions, thecomposition further comprises Methylobacterium strain NLS0017, NLS0020,NLS0021, NLS0042, NLS0064, NLS0066, NLS0089, a derivative thereof, or aMethylobacterium sp. related thereto. In any of the aforementionedembodiments, the plant pathogenic fungi that are inhibited can be intheir anamorphic form, their teleomorphic form, or in both theiranamorphic and teleomorphic forms.

In certain embodiments, the Methylobacterium sp. that inhibit plantpathogenic fungi can be identified by testing newly isolated candidateMethylobacterium sp. for the presence of polymorphic nucleic acid,orthologous gene, or gene sequences that are present in Methylobacteriumsp. provided herein that inhibit certain plant pathogenic fungi.

Various Methylobacterium sp. isolates provided herein are disclosed inTable 1.

TABLE 1 Methylobacterium sp. isolates USDA ARS NLS Origin NRRL No.¹NLS0017 Obtained from a peppermint plant grown NRRL B-50931 in SaintLouis County, Missouri, USA NLS0020 Obtained from a horse nettle plantgrown NRRL B-50930 in Saint Louis County, Missouri, USA NLS0021 Obtainedfrom a lettuce plant grown in NRRL B-50939 Saint Louis Country,Missouri, USA NLS0042 Obtained from a soybean plant grown in NRRLB-50932 Saint Louis Country, Missouri, USA NLS0064 Obtained from a cornplant grown in NRRL B-50938 Saint Louis Country, Missouri, USA NLS0066Obtained from the corn hybrid NRRL B-50940 “MC534” (Masters Choice 3010State Route 146 East Anna, IL 62906) NLS0089 Obtained from a broccoliplant grown NRRL B-50933 in Saint Louis County, Missouri, USA NLS0109Obtained from a Yucca filamentosa plant NRRL B-67340 in Saint LouisCountry, Missouri, USA ¹Deposit number for strain deposited with theAGRICULTURAL RESEARCH SERVICE CULTURE COLLECTION (NRRL) of the NationalCenter for Agricultural Utilization Research, Agricultural ResearchService, U.S. Department of Agriculture, 1815 North University Street,Peoria, Illinois 61604 U.S.A. under the terms of the Budapest Treaty onthe International Recognition of the Deposit of Microorganisms for thePurposes of Patent Procedure. Subject to 37 CFR §1.808(b), allrestrictions imposed by the depositor on the availability to the publicof the deposited material will be irrevocably removed upon the grantingof any patent from this patent application.

Also provided herein are methods for controlling a plant pathogenicfungus that comprise applying any of the aforementioned compositionscomprising the Methylobacterium that are provided herein to a plant or aplant part in an amount that provides for inhibition of infection by theplant pathogenic fungus in the plant, plant part, or a plant obtainedtherefrom relative to infection of a control plant, plant part, or plantobtained therefrom that had not received an application of thecomposition. In certain embodiments, application of the compositionprovides for at least about 40%, at least about 50%, at least about 75%,at least about 85%, or at least about 95% inhibition of a plantpathogenic fungal infection in the plant, plant part, or a plant derivedtherefrom relative to infection of the control plant, plant part, orplant obtained therefrom. In certain embodiments, the plant part isselected from the group consisting of a leaf, a stem, a flower, a root,a tuber, and a seed. In certain embodiments, the method furthercomprises the step of harvesting at least one plant part selected fromthe group consisting of a leaf, a stem, a flower, a root, a tuber, or aseed from the plant or plant part. In certain embodiments of any of theaforementioned methods, the mycotoxin levels in the plant part arereduced by at least 50%, at least 75%, at least 85%, or at least 95%relative to a plant part obtained from the control plant, plant part, orplant obtained therefrom. In certain embodiments of any of theaforementioned methods, the methods further comprise obtaining aprocessed food or feed composition from the plant or plant part. Incertain embodiments of the aforementioned methods, mycotoxin levels inthe processed food or feed composition are reduced by at least 50%, atleast 75%, at least 85%, or at least 95% relative to a processed food orfeed composition obtained from the control plant, plant part, or plantobtained therefrom. In certain embodiments of any of the aforementionedmethods, the composition comprises the Methylobacterium isolate NLS0109,a derivative thereof, or a Methylobacterium related to NLS0109. Incertain embodiments of any of the aforementioned methods, thecomposition further comprises Methylobacterium strain NLS0017, NLS0020,NLS0021, NLS0042, NLS0064, NLS0066, NLS0089, a derivative thereof, or aMethylobacterium sp. related thereto. In certain embodiments of any ofthe aforementioned methods, the composition comprises a Methylobacteriumsp. related to NLS0109 that: (i) has at least one gene encoding a 16SRNA that has at least 97%, 98%, 99%, 99.5%, or 100% sequence identity toSEQ ID NO: 8; (ii) has in its genome one or more polynucleotide markerfragments of at least 50, 60, 100, 120, 180, 200, 240, or 300nucleotides of SEQ ID NOS: 9-11; or (iii) has in its genome one or moremarker fragments comprising a sequence having at least 98%, 99%, or99.5% sequence identity across the entire length of SEQ ID NOS: 9-11. Incertain embodiments of any of the aforementioned methods, thecomposition comprises a Methylobacterium sp. related to NLS0017,NLS0020, NLS0021, NLS0042, NLS0064, NLS0066, or NLS0089 that has atleast one gene encoding a 16S RNA that has at least 97%, 98%, 99%,99.5%, or 100% sequence identity to SEQ ID NO:1, 2, 3, 4, 5, 6, or 7,respectively.

TABLE 2 16S RNA Sequences of Methylobacterium strains SEQ ID NLS NO16S RNA encoding DNA sequence NLS0017 1ggtgatccagccgcaggttcccctacggctaccttgttacgacttcaccccagtcgctgaccctaccgtggtcgcctgcctccttgcggttggcgcagcgccgtcgggtaagaccaactcccatggtgtgacgggcggtgtgtacaaggcccgggaacgtattcaccgtggcatgctgatccacgattactagcgattccgccttcatgcactcgagttgcagagtgcaatccgaactgagacggcttttggggatttgctccagatcgctccttcgcctcccactgtcaccgccattgtagcacgtgtgtagcccatcccgtaagggccatgaggacttgacgtcatccacaccttcctcgcggcttatcaccggcagtctccctagagtgcccaactgaatgatggcaactaaggacgtgggttgcgctcgttgcgggacttaacccaacatctcacgacacgagctgacgacagccatgcagcacctgtgtgcgcgccaccgaagtggaccccaaatctctctgggtaacacgccatgtcaaaggatggtaaggttctgcgcgttgcttcgaattaaaccacatgctccaccgcttgtgcgggcccccgtcaattcctttgagttttaatcttgcgaccgtactccccaggcggaatgctcaaagcgttagctgcgctactgcggtgcaagcaccccaacagctggcattcatcgtttacggcgtggactaccagggtatctaatcctgtttgctccccacgctttcgcgcctcagcgtcagtaatggtccagttggccgccttcgccaccggtgttcttgcgaatatctacgaatttcacctctacactcgcagttccaccaacctctaccatactcaagcgtcccagtatcgaaggccattctgtggttgagccacaggctttcacccccgacttaaaacgccgcctacgcgccctttacgcccagtgattccgagcaacgctagcccccttcgtattaccgcggctgctggcacgaagttagccggggcttattcctccggtaccgtcattatcgtcccggataaaagagctttacaaccctaaggccttcatcactcacgcggcatggctggatcaggcttgcgcccattgtccaatattccccactgctgcctcccgtaggagtctgggccgtgtctcagtcccagtgtggctgatcatcctctcagaccagctactgatcgtcgccttggtaggccgttaccccaccaactagctaatcagacgcgggccgatcttccggcagtaaacctttccccaaaagggcgtatccggtattagccctagtttcccagggttattccgaaccagaaggcacgttcccacgcgttactcacccgtccgccgctgaccccgaaaggcccgctcgacttgcatgtgttaagcctgccgccagcgttcgctctgagccaggatcaaactctc NLS0020 2ggtgatccagccgcaggttcccctacggctaccttgttacgacttcaccccagtcgctgaccctaccgtggtcgcctgcctccttgcggttggcgcagcgccgtcgggtaagaccaactcccatggtgtgacgggcggtgtgtacaaggcccgggaacgtattcaccgtggcatgctgatccacgattactagcgattccgccttcatgcactcgagttgcagagtgcaatccgaactgagacggcttttggggatttgctccagatcgctccttcgcgtcccactgtcaccgccattgtagcacgtgtgtagcccatcccgtaagggccatgaggacttgacgtcatccacaccttcctcgcggcttatcaccggcagtctccctagagtgcccaactgaatgatggcaactaaggacgtgggttgcgctcgttgcgggacttaacccaacatctcacgacacgagctgacgacagccatgcagcacctgtgtgcgcgccaccgaagtggaccccaaatctctctgggtaacacgccatgtcaaaggatggtaaggttctgcgcgttgcttcgaattaaaccacatgctccaccgcttgtgcgggcccccgtcaattcctttgagttttaatcttgcgaccgtactccccaggcggaatgctcaaagcgttagctgcgctactgcggtgcaagcaccccaacagctggcattcatcgtttacggcgtggactaccagggtatctaatcctgtttgctccccacgctttcgcgcctcagcgtcagtaatggtccagttggccgccttcgccaccggtgttcttgcgaatatctacgaatttcacctctacactcgcagttccaccaacctctaccatactcaagcgtcccagtatcgaaggccattctgtggttgagccacaggctttcacccccgacttaaaacgccgcctacgcgccctttacgcccagtgattccgagcaacgctagcccccttcgtattaccgcggctgctggcacgaagttagccggggcttattcctccggtaccgtcattatcgtcccggataaaagagctttacaaccctaaggccttcatcactcacgcggcatggctggatcaggcttgcgcccattgtccaatattccccactgctgcctcccgtaggagtctgggccgtgtctcagtcccagtgtggctgatcatcctctcagaccagctactgatcgtcgccttggtaggccgttaccccaccaactagctaatcagacgcgggccgatcttccggcagtaaacctttccccaaaagggcgtatccggtattagccctagtttcccagggttattccgaaccagaaggcacgttcccacgcgttactcacccgtccgccgctgaccccgaagggcccgctcgacttgcatgtgttaagcctgccgccagcgttcgctctgagccaggatcaaactctc NLS0021 3gagtttgatcctggctcagagcgaacgctggcggcaggcttaacacatgcaagtcgaacgggcttcttcggaagtcagtggcagacgggtgagtaacacgtgggaacgtgcccttcggttcggaataactcagggaaacttgagctaataccggatacgcccttatggggaaaggtttactgccgaaggatcggcccgcgtctgattagcttgttggtggggtaacggcctaccaaggcgacgatcagtagctggtctgagaggatgatcagccacactgggactgagacacggcccagactcctacgggaggcagcagtggggaatattggacaatgggcgcaagcctgatccagccatgccgcgtgagtgatgaaggccttagggttgtaaagctcttttgtccgggacgataatgacggtaccggaagaataagccccggctaacttcgtgccagcagccgcggtaatacgaagggggctagcgttgctcggaatcactgggcgtaaagggcgcgtaggcggccgattaagtcgggggtgaaagcctgtggctcaaccacagaattgccttcgatactggttggcttgagaccggaagaggacagcggaactgcgagtgtagaggtgaaattcgtagatattcgcaagaacaccagtggcgaaggcggctgtctggtccggttctgacgctgaggcgcgaaagcgtggggagcaaacaggattagataccctggtagtccacgccgtaaacgatgaatgccagccgttggtctgcttgcaggtcagtggcgccgctaacgcattaagcattccgcctggggagtacggtcgcaagattaaaactcaaaggaattgacgggggcccgcacaagcggtggagcatgtggtttaattcgaagcaacgcgcagaaccttaccatcccttgacatggcatgttacctcgagagatcggggatcctcttcggaggcgtgcacacaggtgctgcatggctgtcgtcagctcgtgtcgtgagatgttgggttaagtcccgcaacgagcgcaacccacgtccttagttgccatcattcagttgggcactctagggagactgccggtgataagccgcgaggaaggtgtggatgacgtcaagtcctcatggcccttacgggatgggctacacacgtgctacaatggcggtgacagtgggacgcgaaaccgcgaggttgagcaaatccccaaaagccgtctcagttcggattgcactctgcaactcgggtgcatgaaggcggaatcgctagtaatcgtggatcagcacgccacggtgaatacgttcccgggccttgtacacaccgcccgtcacaccatgggagttggtcttacccgacggcgctgcgccaaccgcaagggggcaggcgaccacggtagggtcagcgactggggtgaagtcgtaacaaggtagccgtaggggaacctgcggctggatcacct NLS0042 4ggtgatccagccgcaggttcccctacggctaccttgttacgacttcaccccagtcgctgaccctaccgtggtcgcctgcctccttgcggttggcgcagcgccgtcgggtaagaccaactcccatggtgtgacgggcggtgtgtacaaggcccgggaacgtattcaccgtggcgtgctgatccacgattactagcgattccgccttcatgcacccgagttgcagagtgcaatccgaactgagacggtttttggggatttgctccacctcgcggcttcgcgtcccactgtcaccgccattgtagcacgtgtgtagcccatcccgtaagggccatgaggacttgacgtcatccacaccttcctcgcggcttatcaccggcagtctccctagagtgcccaactgaatgatggcaactaaggacgtgggttgcgctcgttgcgggacttaacccaacatctcacgacacgagctgacgacagccatgcagcacctgtgtgcacgcctccgaagaggatccccgatctctcgaggtaacatgccatgtcaagggatggtaaggttctgcgcgttgcttcgaattaaaccacatgctccaccgcttgtgcgggcccccgtcaattcctttgagttttaatcttgcgaccgtactccccaggcggaatgcttaatgcgttagcggcgccactgacctgcaagcaggccaacggctggcattcatcgtttacggcgtggactaccagggtatctaatcctgtttgctccccacgctttcgcgcctcagcgtcagaaccggaccagacagccgccttcgccactggtgttcttgcgaatatctacgaatttcacctctacactcgcagttccgctgtcctcttccggtctcaagccaaccagtatcgaaggcaattctgtggttgagccacaggctttcacccccgacttaatcggccgcctacgcgccctttacgcccagtgattccgagcaacgctagcccccttcgtattaccgcggctgctggcacgaagttagccggggcttattcttccggtaccgtcattatcgtcccggacaaaagagctttacaaccctaaggccttcatcactcacgcggcatggctggatcaggcttgcgcccattgtccaatattccccactgctgcctcccgtaggagtctgggccgtgtctcagtcccagtgtggctgatcatcctctcagaccagctactgatcgtcgccttggtaggccgttaccccaccaacaagctaatcagacgcgggccgatccttcggcagtaaacctttccccaaaagggcgtatccggtattagctcaagtttccctgagttattccgaaccgaagggtacgttcccacgtgttactcacccgtctgccactgacacccgaaggtgcccgttcgacttgcatgtgttaagcctgccgccagcgttcgctctgagccaggatcaaactctc NLS0064 5ggtgatccagccgcaggttcccctacggctaccttgttacgacttcaccccagtcgctgaccctaccgtggtcgcctgcctccagtcgagcaagctcgatttggttggcgcagcgccgtcgggtaagaccaactcccatggtgtgacgggcggtgtgtacaaggcccgggaacgtattcaccgtggcatgctgatccacgattactagcgattccgccttcatgcacgcgagttgcagcgtgcaatccgaactgagacggcttttggagattggctccgggtcaccccttcgcgtcccactgtcaccgccattgtagcacgtgtgtagcccatcccgtaagggccatgaggacttgacgtcatccacaccttcctcgcggcttatcaccggcagtctccctagagtgcccaaccaaatgatggcaactaaggacgtgggttgcgctcgttgcgggacttaacccaacatctcacgacacgagctgacgacagccatgcagcacctgtgtgcgcgcccccgaaggggacctggaatctctcccagtaacacgccatgtcaaaggatggtaaggttctgcgcgttgcttcgaattaaaccacatgctccaccgcttgtgcgggcccccgtcaattcctttgagttttaatcttgcgaccgtactccccaggcggaatgcttaatgcgttagctgcgctactgcggtgcatgcaccccaacagctagcattcatcgtttacggcgtggactaccagggtatctaatcctgtttgctccccacgctttcgcgcctcagcgtcagtaatggtccagttggccgccttcgccaccggtgttcttgcgaatatctacgaatttcacctctacactcgcagttccaccaacctctaccatactcaagcgtcccagtatcgaaggccattctgtggttgagccacaggctttcacccccgacttaaaacgccgcctacgcgccctttacgcccagtgattccgagcaacgctagcccccttcgtattaccgcggctgctggcacgaagttagccggggcttattcctccggtaccgtcattatcgtcccggagaaaagagctttacaaccctaaggccgtcatcactcacgcggcatggctggatcaggcttgcgcccattgtccaatattccccactgctgcctcccgtaggagtctgggccgtgtctcagtcccagtgtggctgatcatcctctcagaccagctactgatcgtcgccttggtaggccgttaccccaccaacaagctaatcagacgcgggccgatcctccggcagtaaacctttctgccaaagcacgtatccggtattagccctagtttcccagggttatcccagaccggagggcacgttcccacgtgttactcacccgtctgccactcaccttgcggtgcgttcgacttgcatgtgttaagcctgccgccagcgttcgctctgagccaggatcaaactctc NLS0066 6gagtttgatcctggctcagagcgaacgctggcggcaggcttaacacatgcaagtcgaacgcaccgcaaggtgagtggcagacgggtgagtaacacgtgggaacgtgccctccggtctgggataaccctgggaaactagggctaataccggatacgtgctttggcagaaaggtttactgccggaggatcggcccgcgtctgattagcttgttggtggggtaacggcctaccaaggcgacgatcagtagctggtctgagaggatgatcagccacactgggactgagacacggcccagactcctacgggaggcagcagtggggaatattggacaatgggcgcaagcctgatccagccatgccgcgtgagtgatgacggccttagggttgtaaagctcttttctccgggacgataatgacggtaccggaggaataagccccggctaacttcgtgccagcagccgcggtaatacgaagggggctagcgttgctcggaatcactgggcgtaaagggcgcgtaggcggcgttttaagtcgggggtgaaagcctgtggctcaaccacagaatggccttcgatactgggacgcttgagtatggtagaggttggtggaactgcgagtgtagaggtgaaattcgtagatattcgcaagaacaccggtggcgaaggcggccaactggaccattactgacgctgaggcgcgaaagcgtggggagcaaacaggattagataccctggtagtccacgccgtaaacgatgaatgctagctgttggggtgcatgcaccgcagtagcgcagctaacgcattaagcattccgcctggggagtacggtcgcaagattaaaactcaaaggaattgacgggggcccgcacaagcggtggagcatgtggtttaattcgaagcaacgcgcagaaccttaccatcctttgacatggcgtgttactgggagagattccaggtccccttcgggggcgcgcacacaggtgctgcatggctgtcgtcagctcgtgtcgtgagatgttgggttaagtcccgcaacgagcgcaacccacgtccttagttgccatcatttggttgggcactctagggagactgccggtgataagccgcgaggaaggtgtggatgacgtcaagtcctcatggcccttacgggatgggctacacacgtgctacaatggcggtgacagtgggacgcgaaggggtgacccggagccaatctccaaaagccgtctcagttcggattgcacgctgcaactcgcgtgcatgaaggcggaatcgctagtaatcgtggatcagcatgccacggtgaatacgttcccgggccttgtacacaccgcccgtcacaccatgggagttggtcttacccgacggcgctgcgccaaccaaatcgagcttgctcgactggaggcaggcgaccacggtagggtcagcgactggggtgaagtcgtaacaaggtagccgtaggggaacctgcggctggatcacctc NLS0089 7gagtttgatcctggctcagagcgaacgctggcggcaggcttaacacatgcaagtcgaacgggcttcttcggaagtcagtggcagacgggtgagtaacacgtgggaacgtgcccttcggttcggaataactcagggaaacttgagctaataccggatacgcccttacggggaaaggtttactgccgaaggatcggcccgcgtctgattagcttgttggtggggtaacggcctaccaaggcgacgatcagtagctggtctgagaggatgatcagccacactgggactgagacacggcccagactcctacgggaggcagcagtggggaatattggacaatgggcgcaagcctgatccagccatgccgcgtgagtgatgaaggccttagggttgtaaagctcttttgtccgggacgataatgacggtaccggaagaataagccccggctaacttcgtgccagcagccgcggtaatacgaagggggctagcgttgctcggaatcactgggcgtaaagggcgcgtaggcggccgattaagtcgggggtgaaagcctgtggctcaaccacagaattgccttcgatactggttggcttgagaccggaagaggacagcggaactgcgagtgtagaggtgaaattcgtagatattcgcaagaacaccagtggcgaaggcggctgtctggtccggttctgacgctgaggcgcgaaagcgtggggagcaaacaggattagataccctggtagtccacgccgtaaacgatgaatgccagccgttggtctgcttgcaggtcagtggcgccgctaacgcattaagcattccgcctggggagtacggtcgcaagattaaaactcaaaggaattgacgggggcccgcacaagcggtggagcatgtggtttaattcgaagcaacgcgcagaaccttaccatcccttgacatggcatgttacctcgagagatcggggatcctcttcggaggcgtgcacacaggtgctgcatggctgtcgtcagctcgtgtcgtgagatgttgggttaagtcccgcaacgagcgcaacccacgtccttagttgccatcattcagttgggcactctagggagactgccggtgataagccgcgaggaaggtgtggatgacgtcaagtcctcatggcccttacgggatgggctacacacgtgctacaatggcggtgacagtgggacgcgaaaccgcgaggttgagcaaatccccaaaagccgtctcagttcggattgcactctgcaactcgggtgcatgaaggcggaatcgctagtaatcgtggatcagcacgccacggtgaatacgttcccgggccttgtacacaccgcccgtcacaccatgggagttggtcttacccgacggcgctgcgccaaccgcaagggggcaggcgaccacggtagggtcagcgactggggtgaagtcgtaacaaggtagccgtaggggaacctgcggctggatcacct NLS0109 8ggtgatccagccgcaggttcccctacggctaccttgttacgacttcaccccagtcgctgaccctaccgtggtcgcctgctccccttgcgggtcggcgcagcgccgtcgggtaagaccaactcccatggtgtgacgggcggtgtgtacaaggcccgggaacgtattcaccgtggcatgctgatccacgattactagcgattccgccttcatgcactcgagttgcagagtgcaatccgaactgagacggcttttggagatttgcttgccctcgcgggttcgcgtcccactgtcaccgccattgtagcacgtgtgtagcccatcccgtaagggccatgaggacttgacgtcatccacaccttcctcgcggcttatcaccggcagtctccccagagtgcccaactgaatgatggcaactgaggacgtgggttgcgctcgttgcgggacttaacccaacatctcacgacacgagctgacgacagccatgcagcacctgtgtgcgcgctcccgaaggagaccgtggatctctccacgtaacacgccatgtcaaaggatggtaaggttctgcgcgttgcttcgaattaaaccacatgctccaccgcttgtgcgggcccccgtcaattcctttgagttttaatcttgcgaccgtactccccaggcggaatgctcaaagcgttagctgcgccactgagaggcaagccccccaacggctggcattcatcgtttacggcgtggactaccagggtatctaatcctgtttgctccccacgctttcgcgcctcagcgtcagtgtcggaccagttggccgccttcgccaccggtgttcttgcgaatatctacgaatttcacctctacactcgcagttccaccaacctcttccgaactcaagtctcccagtatcgaaggcaattctgtggttgagccacaggctttcacccccgacttaaaagaccgcctacgcgccctttacgcccagtgattccgagcaacgctagcccccttcgtattaccgcggctgctggcacgaagttagccggggcttattcctccggtaccgtcattatcgtcccggataaaagagctttacaaccctaaggccttcatcactcacgcggcatggctggatcaggcttgcgcccattgtccaatattccccactgctgcctcccgtaggagtctgggccgtgtctcagtcccagtgtggctgatcatcctctcagaccagctactgatcgtcgccttggtaggccgttaccccaccaactagctaatcagacgcgggccgatcttccggcagtaaacctttccccaaaagggcgtatccggtattagctcaagtttccctgagttattccgaaccagaaggcacgttcccacgcgttactcacccgtccgccgctgacaccgaagtgcccgctcgacttgcatgtgttaagcctgccgccagcgttcgctctgagccaggatcaaactctc

Also provided are methods of making the compositions useful forcontrolling plant pathogenic fungi that comprise combining aMethylobacterium that inhibit growth of a plant pathogenic fungus withan agriculturally acceptable excipient and/or with an agriculturallyacceptable adjuvant. In certain embodiments of the methods, theMethylobacterium is not M. radiotolerans or M. oryzae. In certainembodiments of any of the aforementioned methods, the compositioncomprises a Methylobacterium sp. related to NLS0109 that has; (i) atleast one gene encoding a 16S RNA that has at least 97%, 98%, 99%,99.5%, or 100% sequence identity to SEQ ID NO: 8; (ii) in its genome oneor more polynucleotide marker fragments of at least 50, 60, 100, 120,180, 200, 240, or 300 nucleotides of SEQ ID NOS: 9-11; or (iii) in itsgenome one or more marker fragments comprising a sequence having atleast 98%, 99%, or 99.5% sequence identity across the entire length ofSEQ ID NOS: 9-11. In certain embodiments of any of the aforementionedmethods, the composition comprises the Methylobacterium isolate NLS0109,a derivative thereof, or a Methylobacterium related to NLS0109. Incertain embodiments of any of the aforementioned methods, thecomposition further comprises Methylobacterium strain NLS0017, NLS0020,NLS0021, NLS0042, NLS0064, NLS0066, NLS0089, a derivative thereof, or aMethylobacterium sp. related thereto. In certain embodiments of themethods, the compositions provide for at least about 25%, at least about50%, or at least about 75% inhibition of plant pathogenic fungal growthin comparison to a control composition that lacks Methylobacterium thatinhibit a plant pathogenic fungus upon exposure to the plant pathogenicfungus. In certain embodiments of the methods, the plant pathogenicfungus is selected from the group consisting of a Blumeria sp., aCercospora sp., a Cochliobolus sp., a Colletotrichum sp., a Diplodiasp., an Exserohilum sp., a Fusarium sp., a Gaeumanomyces sp., aMacrophomina sp., a Magnaporthe sp., a Microdochium sp., a Peronosporasp., a Phakopsora sp., a Phialophora sp., a Phoma sp., a Phymatotrichumsp., a Phytophthora sp., a Pyrenophora sp., a Pyricularia sp, a Pythiumsp., a Rhizoctonia sp., a Sclerophthora, a Sclerospora sp., a Sclerotiumsp., a Sclerotinia sp., a Septoria sp., a Stagonospora sp., aStenocarpella sp. and a Verticillium sp.

In certain embodiments of the methods, the Fusarium sp. is selected fromthe group consisting of Fusarium graminearum, Fusarium verticillioides,Fusarium oxysporum, and Fusarium solani. In certain embodiments of themethods, the Methylobacterium is adhered to a solid substance. Incertain embodiments of the methods, the Methylobacterium adhered to thesolid substance is combined with a liquid to form a composition that isa colloid. In certain embodiments of the methods, the colloid is a gel.In certain embodiments of the methods, the Methylobacterium adhered tothe solid substance is provided by culturing the Methylobacterium in thepresence of the solid substance. In certain embodiments of the methods,the composition comprises an emulsion. In certain embodiments of themethods, the Methylobacterium is provided by culturing theMethylobacterium in an emulsion. In certain embodiments of any of theaforementioned methods, the plant pathogenic fungus is a Fusarium sp.and/or the plant is a cereal plant. In certain embodiments of any of theaforementioned methods, the plant pathogenic fungus is a Fusarium sp.and the plant is a cereal plant selected from the group consisting of arice, wheat, corn, barley, millet, sorghum, oat, and rye plant. Incertain embodiments of any of the aforementioned methods, the plantpathogenic fungus is Fusarium graminearum and the plant is a cerealplant selected from the group consisting of a rice, wheat, corn, barley,millet, sorghum, oat, and rye plant. In any of the aforementionedembodiments, the plant pathogenic fungi that is inhibited can be in itsanamorphic form, its teleomorphic form, or in both its anamorphic andteleomorphic forms.

Methods where Methylobacterium are cultured in biphasic media comprisinga liquid phase and a solid substance have been found to significantlyincrease the resultant yield of Methylobacterium relative to methodswhere the Methylobacterium are cultured in liquid media alone. Incertain embodiments, the methods can comprise growing theMethylobacterium in liquid media with a particulate solid substance thatcan be suspended in the liquid by agitation under conditions thatprovide for Methylobacterium growth. In certain embodiments whereparticulate solid substances are used, at least substantially all of thesolid phase can thus be suspended in the liquid phase upon agitation.Such particulate solid substances can comprise materials that are about1 millimeter or less in length or diameter. In certain embodiments, thedegree of agitation is sufficient to provide for uniform distribution ofthe particulate solid substance in the liquid phase and/or optimallevels of culture aeration. However, in other embodiments providedherein, at least substantially all of the solid phase is not suspendedin the liquid phase, or portions of the solid phase are suspended in theliquid phase and portions of the solid phase are not suspended in theliquid phase. Non-particulate solid substances can be used in certainbiphasic media where the solid phase is not suspended in the liquidphase. Such non-particulate solid substances include, but are notlimited to, materials that are greater than about 1 millimeter in lengthor diameter. Such particulate and non-particulate solid substances alsoinclude, but are not limited to, materials that are porous, fibrous, orotherwise configured to provide for increased surface areas for adherentgrowth of the Methylobacterium. Biphasic media where portions of thesolid phase are suspended in the liquid phase and portions of the solidphase are not suspended in the liquid phase can comprise a mixture ofparticulate and non-particulate solid substances. Such particulate andnon-particulate solid substances used in any of the aforementionedbiphasic media also include, but are not limited to, materials that areporous, fibrous, or otherwise configured to provide for increasedsurface areas for adherent growth of the Methylobacterium. In certainembodiments, the media comprises a colloid formed by a solid and aliquid phase. A colloid comprising a solid and a liquid can bepre-formed and added to liquid media or can be formed in mediacontaining a solid and a liquid. Colloids comprising a solid and aliquid can be formed by subjecting certain solid substances to achemical and/or thermal change. In certain embodiments, the colloid is agel. In certain embodiments, the liquid phase of the media is anemulsion. In certain embodiments, the emulsion comprises an aqueousliquid and a liquid that is not miscible, or only partially miscible, inthe aqueous liquid. Liquids that are not miscible, or only partiallymiscible, in water include, but are not limited to, any of thefollowing: (1) liquids having a miscibility in water that is equal to orless than that of pentanol, hexanol, or heptanol at 25 degrees C.; (2)liquids comprising an alcohol, an aldehyde, a ketone, a fatty acid, aphospholipid, or any combination thereof; (3) alcohols selected from thegroup consisting of aliphatic alcohols containing at least 5 carbons andsterols; (4) an animal oil, microbial oil, synthetic oil, plant oil, orcombination thereof; and/or, (5) a plant oil selected from the groupconsisting of corn, soybean, cotton, peanut, sunflower, olive, flax,coconut, palm, rapeseed, sesame seed, safflower, and combinationsthereof In certain embodiments, the immiscible or partially immiscibleliquid can comprise at least about 0.02% to about 20% of the liquidphase by mass. In certain embodiments, the methods can compriseobtaining a biphasic culture media comprising the liquid, the solid, andMethylobacterium and incubating the culture under conditions thatprovide for growth of the Methylobacterium. Biphasic culture mediascomprising the liquid, the solid, and Methylobacterium can be obtainedby a variety of methods that include, but are not limited to, any of:(a) inoculating a biphasic media comprising the liquid and the solidsubstance with Methylobacterium; (b) inoculating the solid substancewith Methylobacterium and then introducing the solid substancecomprising the Methylobacterium into the liquid media; (c) inoculatingthe solid substance with Methylobacterium, incubating theMethylobacterium on the solid substance, and then introducing the solidsubstance comprising the Methylobacterium into the liquid media; or (d)any combination of (a), (b), or (c). Methods and compositions forgrowing Methylobacterium in biphasic media comprising a liquid and asolid are disclosed in co-assigned US Patent Application Publication No.20130324407, which is incorporated herein by reference in its entirety.

Methods where Methylobacterium are cultured in media comprising anemulsion have also been found to significantly increase the resultantyield of Methylobacterium relative to methods where the Methylobacteriumare cultured in liquid media alone. In certain embodiments, the methodsfor making the compositions provided herein can comprise growing theMethylobacterium in an emulsion under conditions that provide forMethylobacterium growth. Medias comprising the emulsion andMethylobacterium can be obtained by a variety of methods that include,but are not limited to, any of: (a) inoculating a media comprising theemulsion with Methylobacterium; (b) inoculating the aqueous liquid withthe Methylobacterium, introducing the non-aqueous liquid, and mixing toform an emulsion; (c) inoculating the aqueous liquid with theMethylobacterium, introducing the non-aqueous liquid, and mixing to forman emulsion; or (d) any combination of (a), (b), or (c). In certainembodiments, the emulsion comprises an aqueous liquid and a liquid thatis not miscible, or only partially miscible, in the aqueous liquid.Non-aqueous liquids that are not miscible, or only partially miscible,in water include, but are not limited to, any of the following: (1)liquids having a miscibility in water that is equal to or less than thatof n-pentanol, n-hexanol, or n-heptanol at 25 degrees C.; (2) liquidscomprising an alcohol, an aldehyde, a ketone, a fatty acid, aphospholipid, or any combination thereof; (3) alcohols selected from thegroup consisting of aliphatic alcohols containing at least 5, 6, or 7carbons and sterols; (4) an animal oil, microbial oil, synthetic oil,plant oil, or combination thereof; and/or, (5) a plant oil selected fromthe group consisting of corn, soybean, cotton, peanut, sunflower, olive,flax, coconut, palm, rapeseed, sesame seed, safflower, and combinationsthereof. In certain embodiments, the immiscible or partially immisciblenon-aqueous liquid can comprise at least about 0.02% to about 20% of theemulsion by mass. In certain embodiments, the immiscible or partiallyimmiscible non-aqueous liquid can comprise at least about any of about0.05%, 0.1%, 0.5%, or 1% to about 3%, 5%, 10%, or 20% of the emulsion bymass. Methods and compositions for growing Methylobacterium in mediacomprising an emulsion are disclosed in co-assigned U.S. patentapplication Ser. No. 14/894,568, filed May 30, 2014, which isincorporated herein by reference in its entirety.

In certain embodiments, the fermentation broth, fermentation brothproduct, or compositions that comprise Methylobacterium that inhibitplant pathogenic fungi can further comprise one or more introducedmicroorganisms of pre-determined identity other than Methylobacterium.Other microorganisms that can be added include, but are not limited to,microorganisms that are biopesticidal or provide some other benefit whenapplied to a plant or plant part. Biopesticidal or otherwise beneficialmicroorganisms thus include, but are not limited to, various Bacillussp., Pseudomonas sp., Coniothyrium sp., Pantoea sp., Streptomyces sp.,and Trichoderma sp., as well as bioinoculants for improved nitrogenfixation, including rhizobia species such as from Rhizobium orBradyrhizobium, Microbial biopesticides can be a bacterium, fungus,virus, or protozoan. Particularly useful biopesticidal microorganismsinclude various Bacillus subtilis, Bacillus thuringiensis, Bacilluspumilis, Pseudomonas syringae, Trichoderma harzianum, Trichodermavixens, and Streptomyces lydicus strains. Other microorganisms that areadded can be genetically engineered or isolates that are available aspure cultures. In certain embodiments, it is anticipated that thebacterial or fungal microorganism can be provided in the fermentationbroth, fermentation broth product, or composition in the form of aspore.

In certain embodiments, the liquid culture medium is prepared frominexpensive and readily available components, including, but not limitedto, inorganic salts such as potassium phosphate, magnesium sulfate andthe like, carbon sources such as glycerol, methanol, glutamic acid,aspartic acid, succinic acid and the like, and amino acid blends such aspeptone, tryptone, and the like. Examples of liquid media that can beused include, but are not limited to, ammonium mineral salts (AMS)medium (Whittenbury et al., 1970), Vogel-Bonner (VB) minimal culturemedium (Vogel and Bonner, 1956), and LB broth (“Luria—Bertani Broth”).

Fermentation products and compositions with a mono- or co-culture ofMethylobacterium that inhibit plant pathogenic fungi at a titer ofgreater than about 5×10⁷ colony-forming units per milliliter, at a titerof greater than about 1×10⁸ colony-forming units per milliliter, at atiter of greater than about 5×10⁸ colony-forming units per milliliter,at a titer of greater than about 1×10⁹ colony-forming units permilliliter, at a titer of greater than about 1×10¹⁰ colony-forming unitsper milliliter, at a titer of at least about 3×10¹⁰ colony-forming unitsper milliliter are provided herein. In certain embodiments, fermentationproducts and compositions provided herein can comprise Methylobacteriumthat inhibit plant pathogenic fungi at a titer of at least about 5×10⁷,1×10⁸ , or 5×10⁸ colony-forming units per milliliter to at least about3×10¹⁰ colony-forming units per milliliter, at least about 5×10⁸colony-forming units per milliliter to at least about 4×10¹⁰colony-forming units per milliliter, or at least about 5×10⁸colony-forming units per milliliter to at least about 6×10¹⁰colony-forming units per milliliter. In certain embodiments,fermentation products and compositions provided herein can compriseMethylobacterium that inhibit plant pathogenic fungi at a titer of atleast about 1×10⁹ colony-forming units per milliliter to at least about3×10¹⁰ colony-forming units per milliliter, at least about 1×10⁹colony-forming units per milliliter to at least about 4×10¹⁰colony-forming units per milliliter, or at least about 1×10⁹colony-forming units per milliliter to at least about 6×10¹⁰colony-forming units per milliliter. In certain embodiments,fermentation products and compositions provided herein will compriseMethylobacterium that inhibit plant pathogenic fungi at a titer of atleast about 1×10¹⁰ colony-forming units per milliliter to at least about3×10¹⁰ colony-forming units per milliliter, at least about 1×10¹⁰colony-forming units per milliliter to at least about 4×10¹⁰colony-forming units per milliliter, or at least about 1×10¹⁰colony-forming units per milliliter to at least about 6×10¹⁰colony-forming units per milliliter. In certain embodiments,fermentation products and compositions provided herein will compriseMethylobacterium that inhibit plant pathogenic fungi at a titer of, atleast about 3×10¹⁰ colony-forming units per milliliter to at least about4×10¹⁰ colony-forming units per milliliter, or at least about 3×10¹⁰colony-forming units per milliliter to at least about 6×10¹⁰colony-forming units per milliliter. In any of the aforementionedfermentation products or compositions, the indicated concentrations canbe fungal inhibitory concentrations. In any of the aforementionedfermentation products or compositions, the fermentation products orcompositions can be essentially free of contaminating microorganisms,can comprise Methylobacterium that are adhered to and/or associated withmaterials that the Methylobacterium are not are adhered to and/orassociated with in nature, or any combination thereof.

Fermentation products and compositions with Methylobacterium thatinhibit plant pathogenic fungi at a titer of greater than about 5×10⁷,1×10⁸, or 5×10⁸ colony-forming units per gram, at a titer of greaterthan about 1×10⁹ colony-forming units per gram, at a titer of greaterthan about 1×10¹⁰ colony-forming units per gram, at a titer of at leastabout 3×10¹⁰ colony-forming units per gram are provided herein. Incertain embodiments, fermentation products and compositions providedherein can comprise Methylobacterium that inhibit plant pathogenic fungiat a titer of at least about 5×10⁷, 1×10⁸ , or 5×10⁸ colony-formingunits per gram to at least about 3×10¹⁰ colony-forming units per gram,at least about 5×10⁷ , 1×10⁸ , or 5×10⁸ colony-forming units per gram toat least about 4×10¹⁰ colony-forming units per gram, or at least about5×10⁷ , 1×10⁸ , or 5×10⁸ colony-forming units per gram to at least about6×10¹⁰ colony-forming units per gram. In certain embodiments,fermentation products and compositions provided herein can compriseMethylobacterium that inhibit plant pathogenic fungi at a titer of atleast about 1×10⁹ colony-forming units per gram to at least about 3×10¹⁰colony-forming units per gram, at least about 1×10⁹ colony-forming unitsper gram to at least about 4×10¹⁰ colony-forming units per gram, or atleast about 1×10⁹ colony-forming units per gram to at least about 6×10¹⁰colony-forming units per gram. In certain embodiments, fermentationproducts and compositions provided herein will comprise Methylobacteriumthat inhibit plant pathogenic fungi at a titer of at least about 1×10¹⁰colony-forming units per gram to at least about 3×10¹⁰ colony-formingunits per gram, at least about 1×10¹⁰ colony-forming units per gram toat least about 4×10¹⁰ colony-forming units per gram, or at least about1×10¹⁰ colony-forming units per gram to at least about 6×10¹⁰colony-forming units per gram. In certain embodiments, fermentationproducts and compositions provided herein will comprise Methylobacteriumthat inhibit plant pathogenic fungi at a titer of, at least about 3×10¹⁰colony-forming units per gram to at least about 4×10¹⁰ colony-formingunits per gram, or at least about 3×10¹⁰ colony-forming units per gramto at least about 6×10¹⁰, 1×10¹¹, 1×10¹², 1×10¹³, or 5×10¹³colony-forming units per gram. In any of the aforementioned fermentationproducts or compositions, the fermentation or composition can comprise amono- or co-culture of Methylobacterium that is adhered to a solidsubstance. In any of the aforementioned fermentation products orcompositions, the indicated concentrations can be fungal inhibitoryconcentrations. In any of the aforementioned fermentation products orcompositions, the indicated concentrations can be fungal inhibitoryconcentrations. In any of the aforementioned fermentation products orcompositions, the fermentation products or compositions can beessentially free of contaminating microorganisms, can compriseMethylobacterium that are adhered to and/or associated with materialsthat the Methylobacterium are not are adhered to and/or associated within nature, or any combination thereof.

Methylobacterium that inhibit plant pathogenic fungi can be obtained asfermentation products can be used to make various compositions usefulfor treating plants or plant parts to inhibit infection by plantpathogenic fungi. Alternatively, compositions provided herein comprisingsolid substances with Methylobacterium that inhibit plant pathogenicfungi or adherent Methylobacterium that inhibit plant pathogenic fungican be used to treat plants or plant parts. Plants, plant parts, and, inparticular, plant seeds that have been at least partially coated withthe fermentation broth products or compositions comprisingMethylobacterium that inhibit plant pathogenic fungi are thus provided.Also provided are processed plant products that contain the fermentationbroth products or compositions with Methylobacterium that inhibit plantpathogenic fungi or adherent Methylobacterium that inhibit plantpathogenic fungi. Methylobacterium that inhibit plant pathogenic fungican be used to make various compositions that are particularly usefulfor treating plant seeds. Seeds that have been at least partially coatedwith the fermentation broth products or compositions are thus provided.Also provided are processed seed products, including, but not limitedto, meal, flour, feed, and flakes that contain the fermentation brothproducts or compositions provided herein. In certain embodiments, theprocessed plant product will be non-regenerable (i.e. will be incapableof developing into a plant). In certain embodiments, the fermentationproduct or composition that at least partially coats the plant, plantpart, or plant seed or that is contained in the processed plant, plantpart, or seed product comprises a Methylobacterium that inhibit plantpathogenic fungi that can be readily identified by comparing a treatedand an untreated plant, plant part, plant seed, or processed productthereof. In certain embodiments, the identification is performed bydetermining if the fermentation product or composition that at leastpartially coats the plant, plant part, or plant seed or that iscontained in the processed plant, plant part, or seed product comprisesa Methylobacterium sp. related to NLS0109 that: (i) has at least onegene encoding a 16S RNA that has at least 97%, 98%, 99%, 99.5%, or 100%sequence identity to SEQ ID NO: 8; (ii) has in its genome one or morepolynucleotide marker fragments of at least 50, 60, 100, 120, 180, 200,240, or 300 nucleotides of SEQ ID NOS: 9-11; or (iii) has in its genomeone or more marker fragments comprising a sequence having at least 98%,99%, or 99.5% sequence identity across the entire length of SEQ ID NOS:9-11. Similar 16S RNA sequence analyses can be conducted to determine ifthe fermentation product or composition that at least partially coatsthe plant, plant part, or plant seed or that is contained in theprocessed plant, plant part, or seed product further comprises NLS0017,NLS0020, NLS0021, NLS0042, NLS0064, NLS0066, NLS0089, a derivativethereof, or a Methylobacterium sp. related thereto.

Compositions useful for treating plants or plant parts that compriseMethylobacteriumthat inhibit plant pathogenic fungi or a solid substancewith adherent Methylobacterium that inhibit plant pathogenic fungi,emulsions containing the Methylobacterium that inhibit plant pathogenicfungi or combinations thereof can also comprise an agriculturallyacceptable adjuvant or an agriculturally acceptable excipient. Anagriculturally acceptable adjuvant or an agriculturally acceptableexcipient is typically an ingredient that does not cause unduephytotoxicity or other adverse effects when exposed to a plant or plantpart. In certain embodiments, the solid substance can itself be anagriculturally acceptable adjuvant or an agriculturally acceptableexcipient so long as it is not bacteriocidal or bacteriostatic to theMethylobacterium. In other embodiments, the composition furthercomprises at least one of an agriculturally acceptable adjuvant or anagriculturally acceptable excipient. Any of the aforementionedcompositions can also further comprise a pesticide. Pesticides used inthe composition include, but are not limited to, an insecticide, afungicide, a nematocide, and a bacteriocide. In certain embodiments, thepesticide used in the composition is a pesticide that does notsubstantially inhibit growth of the Methylobacterium. AsMethylobacterium are gram negative bacteria, suitable bacteriocides usedin the compositions can include, but are not limited to, bacteriocidesthat exhibit activity against gram positive bacteria but not gramnegative bacteria. Compositions provided herein can also comprise abacteriostatic agent that does not substantially inhibit growth of theMethylobacterium. Bacteriostatic agents suitable for use in compositionsprovided herein include, but are not limited to, those that exhibitactivity against gram positive bacteria but not gram negative bacteria.Any of the aforementioned compositions can also be an essentially dryproduct (i.e. having about 5% or less water content), a mixture of thecomposition with an emulsion, or a suspension.

Agriculturally acceptable adjuvants used in the compositions thatcomprise Methylobacterium that inhibit plant pathogenic fungi, emulsionscontaining the Methylobacterium that inhibit plant pathogenic fungi, orcombinations thereof include, but are not limited to, components thatenhance product efficacy and/or products that enhance ease of productapplication. Adjuvants that enhance product efficacy can include variouswetters/spreaders that promote adhesion to and spreading of thecomposition on plant parts, stickers that promote adhesion to the plantpart, penetrants, extenders, and humectants that increase the density ordrying time of sprayed compositions. Wetters/spreaders used in thecompositions can include, but are not limited to, non-ionic surfactants,anionic surfactants, cationic surfactants, amphoteric surfactants,organo-silicate surfactants, and/or acidified surfactants. Stickers usedin the compositions can include, but are not limited to, latex-basedsubstances, terpene/pinolene, and pyrrolidone-based substances.Penetrants can include mineral oil, vegetable oil, esterified vegetableoil, organo-silicate surfactants, and acidified surfactants. Extendersused in the compositions can include, but are not limited to, ammoniumsulphate, or menthene-based substances. Humectants used in thecompositions can include, but are not limited to, glycerol, propyleneglycol, and diethyl glycol. Adjuvants that improve ease of productapplication include, but are not limited to, acidifying/bufferingagents, anti-foaming/de-foaming agents, compatibility agents,drift-reducing agents, dyes, and water conditioners.Anti-foaming/de-foaming agents used in the compositions can include, butare not limited to, dimethopolysiloxane. Compatibility agents used inthe compositions can include, but are not limited to, ammonium sulphate.Drift-reducing agents used in the compositions can include, but are notlimited to, polyacrylamides, and polysaccharides. Water conditionersused in the compositions can include, but are not limited to, ammoniumsulphate.

Methods of treating plants and/or plant parts with the fermentationbroths, fermentation broth products, and compositions comprisingMethylobacterium that inhibit plant pathogenic fungi, or combinationsthereof are also provided herein. Treated plants, and treated plantparts obtained therefrom, include, but are not limited to, corn,Brassica sp. (e.g., B. napus, B. rapa, B. juncea), alfalfa, rice, rye,sorghum, millet (e.g., pearl millet (Pennisetum glaucum)), proso millet(Panicum miliaceum), foxtail millet (Setaria italica), finger millet(Eleusine coracana), sunflower, safflower, soybean, tobacco, potato,peanuts, cotton, sweet potato (Ipomoea batatus), cassava, coffee,coconut, pineapple, citrus trees, cocoa, tea, banana, avocado, fig,guava, mango, olive, papaya, cashew, macadamia, almond, sugar beets,sugarcane, oats, barley, tomatoes, lettuce, green beans, lima beans,peas, cucurbits such as cucumber, cantaloupe, and musk melon,ornamentals, and conifers. Plant parts that are treated include, but arenot limited to, leaves, stems, flowers, roots, seeds, fruit, tubers,coleoptiles, and the like. Ornamental plants and plant parts that can betreated include, but are not limited to azalea, hydrangea, hibiscus,roses, tulips, daffodils, petunias, carnation, poinsettia, andchrysanthemum. Conifer plants and plant parts that can be treatedinclude, but are not limited to, pines such as loblolly pine, slashpine, ponderosa pine, lodge pole pine, and Monterey pine; Douglas-fir;Western hemlock; Sitka spruce; redwood; true firs such as silver fir andbalsam fir; and cedars such as Western red cedar and Alaskayellow-cedar. Turfgrass plants and plant parts that can be treatedinclude, but are not limited to, annual bluegrass, annual ryegrass,Canada bluegrass, fescue, bentgrass, wheatgrass, Kentucky bluegrass,orchard grass, ryegrass, redtop, Bermuda grass, St. Augustine grass, andzoysia grass. In certain embodiments, the treated plant or plant part isa cereal plant or plant part selected from the group consisting of arice, wheat, corn, barley, millet, sorghum, oat, and rye plant or plantpart. Seeds or other propagules of any of the aforementioned plants canbe treated with the fermentation broths, fermentation broth products,fermentation products, and/or compositions provided herein.

In certain embodiments, plants and/or plant parts are treated byapplying the fermentation broths, fermentation broth products,fermentation products, and compositions that comprise Methylobacteriumthat inhibit plant pathogenic fungi, or combinations thereof as a spray.Such spray applications include, but are not limited to, treatments of asingle plant part or any combination of plant parts. Spraying can beachieved with any device that will distribute the fermentation broths,fermentation broth products, fermentation products, and compositions tothe plant and/or plant part(s). Useful spray devices include a boomsprayer, a hand or backpack sprayer, crop dusters (i.e. aerialspraying), and the like. Spraying devices and or methods providing forapplication of the fermentation broths, fermentation broth products,fermentation products, and compositions to either one or both of theadaxial surface and/or abaxial surface can also be used. Plants and/orplant parts that are at least partially coated with any of a biphasicfermentation broth, a fermentation broth product, fermentation product,or compositions that comprise a solid substance with Methylobacteriumthat inhibit plant pathogenic fungi adhered thereto are also providedherein. Also provided herein are processed plant products that comprisea solid substance with Methylobacterium that inhibit plant pathogenicfungi adhered thereto.

In certain embodiments, seeds are treated by exposing the seeds to thefermentation broths, fermentation broth products, fermentation products,and compositions that comprise Methylobacterium that inhibit plantpathogenic fungi, or combinations thereof. Seeds can be treated with thefermentation broths, fermentation broth products, and compositionsprovided herein by methods including, but not limited to, imbibition,coating, spraying, and the like. Seed treatments can be effected withboth continuous and/or a batch seed treaters. In certain embodiments,the coated seeds can be prepared by slurrying seeds with a coatingcomposition containing a fermentation broth, fermentation broth product,or compositions that comprise the solid substance with Methylobacteriumthat inhibit plant pathogenic fungi and air drying the resultingproduct. Air drying can be accomplished at any temperature that is notdeleterious to the seed or the Methylobacterium, but will typically notbe greater than 30 degrees Centigrade. The proportion of coating thatcomprises a solid substance and Methylobacterium that inhibit plantpathogenic fungi includes, but is not limited to, a range of 0.1 to 25%by weight of the seed, 0.5 to 5% by weight of the seed, and 0.5 to 2.5%by weight of seed. In certain embodiments, a solid substance used in theseed coating or treatment will have Methylobacterium that inhibit plantpathogenic fungi adhered thereon. In certain embodiments, a solidsubstance used in the seed coating or treatment will be associated withMethylobacterium that inhibit plant pathogenic fungi and will be afermentation broth, fermentation broth product, or composition obtainedby the methods provided herein. Various seed treatment compositions andmethods for seed treatment disclosed in U.S. Pat. Nos. 5,106,648;5,512,069; and 8,181,388 are incorporated herein by reference in theirentireties and can be adapted for use with fermentation products orcompositions provided herein. In certain embodiments, the compositionused to treat the seed can contain agriculturally acceptable excipientsthat include, but are not limited to, woodflours, clays, activatedcarbon, diatomaceous earth, fine-grain inorganic solids, calciumcarbonate and the like. Clays and inorganic solids that can be used withthe fermentation broths, fermentation broth products, or compositionsprovided herein include, but are not limited to, calcium bentonite,kaolin, china clay, talc, perlite, mica, vermiculite, silicas, quartzpowder, montmorillonite and mixtures thereof. Agriculturally acceptableadjuvants that promote sticking to the seed that can be used include,but are not limited to, polyvinyl acetates, polyvinyl acetatecopolymers, hydrolyzed polyvinyl acetates, polyvinylpyrrolidone-vinylacetate copolymer, polyvinyl alcohols, polyvinyl alcohol copolymers,polyvinyl methyl ether, polyvinyl methyl ether-maleic anhydridecopolymer, waxes, latex polymers, celluloses including ethylcellulosesand methylcelluloses, hydroxy methylcelluloses, hydroxypropylcellulose,hydroxymethylpropylcelluloses, polyvinyl pyrrolidones, alginates,dextrins, malto-dextrins, polysaccharides, fats, oils, proteins, karayagum, jaguar gum, tragacanth gum, polysaccharide gums, mucilage, gumarabics, shellacs, vinylidene chloride polymers and copolymers,soybean-based protein polymers and copolymers, lignosulfonates, acryliccopolymers, starches, polyvinylacrylates, zeins, gelatin,carboxymethylcellulose, chitosan, polyethylene oxide, acrylamidepolymers and copolymers, polyhydroxyethyl acrylate, methylacrylamidemonomers, alginate, ethylcellulose, polychloroprene and syrups ormixtures thereof. Other useful agriculturally acceptable adjuvants thatcan promote coating include, but are not limited to, polymers andcopolymers of vinyl acetate, polyvinylpyrrolidone-vinyl acetatecopolymer and water-soluble waxes. Various surfactants, dispersants,anticaking-agents, foam-control agents, and dyes disclosed herein and inU.S. Pat. No. 8,181,388 can be adapted for use with a fermentationproducts or compositions provided herein.

Provided herein are compositions that comprise Methylobacterium thatinhibit plant pathogenic fungi and that provide control of plantpathogenic fungal infections of plants, plant parts, and plants obtainedtherefrom relative to untreated plants, plant parts, and plants obtainedtherefrom that have not been exposed to the compositions. In certainembodiments, plant parts, including, but not limited to, a seed, a leaf,a fruit, a stem, a root, a tuber, or a coleoptile can be treated withthe compositions provided herein to control fungal disease. Treatmentsor applications can include, but are not limited to, spraying, coating,partially coating, immersing, and/or imbibing the plant or plant partswith the compositions provided herein. In certain embodiments, a seed, aleaf, a fruit, a stem, a root, a tuber, or a coleoptile can be immersedand/or imbibed with a liquid, semi-liquid, emulsion, or slurry of acomposition provided herein. Such seed immersion or imbibition can besufficient to provide for fungal disease inhibition in a plant or plantpart in comparison to an untreated plant or plant part. Such fungaldisease inhibition includes, but is not limited to decreases in fungalgrowth and/or the adverse effects of fungal growth relative to untreatedplants. In certain embodiments, plant seeds can be immersed and/orimbibed for at least 1, 2, 3, 4, 5, or 6 hours. Such immersion and/orimbibition can, in certain embodiments, be conducted at temperaturesthat are not deleterious to the plant seed or the Methylobacterium. Incertain embodiments, the seeds can be treated at about 15 to about 30degrees Centigrade or at about 20 to about 25 degrees Centigrade. Incertain embodiments, seed imbibition and/or immersion can be performedwith gentle agitation.

Amounts of the compositions that comprise Methylobacterium that inhibitplant pathogenic fungi that are sufficient to provide for an inhibitionof fungal infection of a plant or plant part can thus be determined bymeasuring any or all fungal growth and/or the adverse effects of fungalgrowth in treated plants or plant parts relative to untreated plants orplant parts. Adverse effects of fungal growth in a plant that can bemeasured include any type of plant tissue damage or necrosis, any typeof plant yield reduction, any reduction in the value of the crop plantproduct, and/or production of undesirable fungal metabolites or fungalgrowth by-products including, but not limited to, mycotoxins. Mycotoxinscomprise a number of toxic molecules produced by fungal species,including, but not limited to, polyketides (including aflatoxins,demethylsterigmatocystin, O-methylsterigmatocystin etc.), fumonisins,alperisins (e.g., A₁, A₂, B₁, B₂), sphingofungins (A, B, C and D),trichothecenes, fumifungins, and the like. Methods of quantitatingmycotoxin levels are widely documented. Moreover, commercial kits formeasurement of the mycotoxins such as aflatoxin, fumonisin,deoxynivalenol, and zearalenone are also available (VICAM, Watertown,Mass., USA).

Compositions provided herein comprising Methylobacterium that inhibitplant pathogenic fungi are therefore expected to be useful in inhibitingfungal growth and/or infection in a wide variety of plant pathogenicfungi, including, but not limited to the anamorphic and/or teleomorphicstages of those phytopathogenic fungi in the following genera andspecies: Blumeria (Blumeria graminis f. sp. tritici), Cercospora(Cercospora kikuchii; Cercospora sojina; Cercospora zeae-maydis);Cochliobolus (Colchliobolus maydis; Cochliobolus heterostrophus;Cochliobolus carbonum); Colletotrichum (Colletotrichum lindemuthianum;Colletotrichum graminicola; Colletotrichum cereale); Diplodia (Diplodiamaydis); Exserohilum (Exserohilum turcicum); Fusarium (Fusarium nivale;Fusarium oxysporum; Fusarium graminearum; Fusarium culmorum; Fusariumsolani; Fusarium moniliforme; Fusarium virguliforme); Macrophomina(Macrophomina phaseolina); Magnaporthe (Magnaporthe oryzae; Magnaporthegrisea); Phakopsora (Phakopsora pachyrhizi); Phialopora (Phialophoragregata); Phymatotrichum (Phymatotrichum omnivorum); Phytophthora(Phytophthora cinnamomi; Phytophthora cactorum; Phytophthora phaseoli;Phytophthora parasitica; Phytophthora citrophthora; Phytophthoramegasperma f. sp. sojae; Phytophthora infestans); Puccinia (Pucciniasorghi; Puccinia striiformis; Puccinia graminis f sp. tritici; Pucciniaasparagi; Puccinia recondita; Puccinia arachidis; Puccinia coronata);Pythium (Pythium aphanidermatum; Pythium ultimum; Pythium sylvaticum;Pythium torulosum; Pythium lutarium; Pythium oopapillum); Pyrenophora(Pyrenophora tritici-repentis); Rhizoctonia (Rhizoctonia solani;Rhizoctonia cerealis); Sclerotium (Sclerotium rolfsii); Sclerotinia(Sclerotinia sclerotiorum; Sclerotinia homoeocarpa); Septoria (Septorialycopersici; Septoria glycines; Septoria nodorum; Septoria tritici);Setosphaeria (Setosphaeria turcica); Stagonospora (Stagonosporanodorum); Verticillium (Verticillium dahliae; Verticillium albo-atrum).Compositions provided herein comprising Methylobacterium that inhibitplant pathogenic fungi are also expected to be useful in inhibitingfungal growth and/or infection by Fusarium graminearum, Fusariumverticillioides, Fusarium virguliforme, and/or Fusarium proliferatum.Compositions provided herein comprising Methylobacterium that inhibitfungal growth and/or infection by Fusarium graminearum, Fusariumverticillioides, Fusarium virguliforme, and/or Fusarium proliferatum canbe used to control infections of cereal plants infected by these fungi.Infections of cereal plants selected from the group consisting of arice, wheat, corn, barley, millet, sorghum, oat, and rye plants byFusarium sp can be controlled by the compositions provided herein. Inany of the aforementioned embodiments, the plant pathogenic fungus thatis inhibited can be in its anamorphic form, its teleomorphic form, or inboth its anamorphic and teleomorphic forms. Certain Methylobacteriumisolates or combinations of isolates can also be used to inhibit certainplant pathogenic fungi in certain crops as disclosed in Table 3. Incertain embodiments where a combination of isolates are used (e.g.,NLS0109, a derivative thereof, or a Methylobacterium sp. related theretoand at least one additional Methylobacterium strain selected from thegroup consisting of NLS0017, NLS0020, NLS0021, NLS0042, NLS0064,NLS0066, NLS0089, a derivative thereof, and a Methylobacterium sp.related thereto), the isolates can be applied either simultaneously orsequentially. In certain embodiments where a combination of isolates areused (e.g., or NLS0109, a derivative thereof, or a Methylobacterium sp.related thereto and at least one additional Methylobacterium strainselected from the group consisting of NLS0017, NLS0020, NLS0021,NLS0042, NLS0064, NLS0066, NLS0089, a derivative thereof, and aMethylobacterium sp. related thereto), the isolates can be applied ineither the same mode(s) (e.g., via a seed treatment, a foliarapplication, or in furrow) or by distinct modes.

TABLE 3 Methylobacterium isolates and combinations of isolates for usein compositions and methods for controlling certain plant pathogenicfungi in certain crops (including derivatives of the Methylobacteriumisolates or Methylobacterium spp. related thereto) NLS Isolate(s) CropPathogen Disease Common Name(s) Mode(s) of Application NLS0109 WheatFusarium Fusarium head blight; Seed treatment; graminearum seedlingblight foliar; or both Pythium spp Pythium root rot Septoria tritici;Septoria/Stagonospora Stagonospora nodorum blotch Rhizoctonia solaniRhizoctonia root rot Fusarium spp. Fusarium root, crown, and foot rotMagnaporthe grisea Head blast Pyrenophora tritici- Tan Spot repentisMicrodochium nivale Snow mold Blumeria graminis f. Powdery mildew sp.tritici NLS0109 Corn Cercospora zeae- Gray leaf spot Seed treatment;maydis In-furrow; foliar; or Colletotrichum Anthracnose (foliar bothin-furrow and graminicola and stalk rot) foliar Fusarium Fusarium stalkrot; ear graminearum rot Fusarium Fusarium ear rot verticillioidesRhizoctonia solani Rhizoctonia crown and root rot Stenocarpella maydisDiplodia stalk and ear rot Pythium spp. Pythium root rot SclerosporaDowny mildew; Crazy graminicola; top Sclerophthora macrospora NLS0109Soybean Cercospora sojina Frogeye leaf spot Seed treatment; Cercosporiakikuchii Purple leaf blotch and in-furrow; foliar; or seed stain anycombination Septoria glycines Brown spot thereof Pythium spp. Pythiumroot rot Fusarium spp. Fusarium seed rot, blight/wilt, root rot, and podand collar rot Sclerotinia White mold sclerotiorum Fusarium virguliformSudden death syndrome Rhizoctonia solani Rhizoctonia damping off androot rot Peronospora Downy mildew manshurica Phytophthora sojae Root andstem rot NLS0109 + Wheat Fusarium Fusarium head blight; Seed treatment;NLS0089 graminearum seedling blight in-furrow; foliar; or Septoriatritici; Septoria/Stagonospora any combination Stagonospora nodorumblotch thereof where Pythium spp. Pythium root rot isolates are usedRhizoctonia solani Rhizoctonia root rot alone or in Fusarium spp.Fusarium root, crown, combination for and foot rot each treatment.Blumeria graminis f. Powdery mildew sp. tritici Magnaporthe grisea Headblast Pyrenophora tritici- Tan Spot repentis Microdochium nivale Snowmold NLS0109 + Soybean Sclerotinia White mold Seed treatment; NLS0089sclerotiorum in-furrow; foliar; or Cercospora sojina Frogeye leaf spotany combination Cercospora kikuchii Purple leaf blotch and thereof,where seed stain isolates are used Fusarium spp. Fusarium seed rot,alone or in blight/wilt, root rot, combination for and pod and collarrot any treatment. Septoria glycines Brown spot Pythium spp. Pythiumroot rot Peronospora Downy mildew manshurica Rhizoctonia solaniRhizoctonia damping off and root rot Phytophthora sojae Root and stemrot Fusarium virguliform Sudden death syndrome NLS0109 + Corn Pythiumspp. Pythium root and stalk In-furrow; foliar; or NLS0089 rot anycombination Stenocarpella maydis Diplodia stalk and ear thereof, whererot isolates are used Sclerospora Downy mildew; Crazy alone or ingraminicola; top combination for Sclerophthora any treatment. macrosporaRhizoctonia solani Rhizoctonia crown and root rot Fusarium spp. Fusariumroot and stalk rot Colletotrichum Anthracnose leaf blight graminicolaand stalk rot Cercospora zeae- Gray leaf spot maydis NLS0020 + WheatFusarium Fusarium head blight; Seed treatment; NLS0109 graminearumseedling blight in-furrow; foliar; or Septoria tritici;Septoria/Stagonospora any combination Stagonospora nodorum blotchthereof, where Pythium spp. Pythium root rot isolates are usedRhizoctonia solani Rhizoctonia root rot alone or in Fusarium spp.Fusarium root, crown, combination for and foot rot any treatment,Blumeria graminis f. Powdery mildew including NLS0020 sp. triticiapplied in-furrow Magnaporthe grisea Head blast and NLS0109 Pyrenophoratritici- Tan spot applied foliar. repentis Microdochium nivale Snow moldNLS0020 + Soybean Sclerotinia White mold Seed treatment; NLS0109sclerotiorum in-furrow; foliar; or Cercospora sojina Frogeye leaf spotany combination Cercospora kikuchii Purple leaf blotch and thereof,where seed stain isolates are used Septoria glycines Brown spot alone orin Pythium spp. Pythium root rot combination for Fusarium spp. Fusariumseed rot, any treatment, blight/wilt, root rot, including NLS0020 andpod and collar rot applied in-furrow Peronospora Downy mildew andNLS0109 manshurica applied foliar. Rhizoctonia solani Rhizoctoniadamping off and root rot Phytophthora sojae Root and stem rot Fusariumvirguliform Sudden death syndrome NLS0020 + Corn Cercospora zeae- Grayleaf spot Seed treatment; NLS0109 maydis in-furrow; foliar; orColletotrichum Anthracnose (foliar any combination graminicola and stalkrot) thereof, where Fusarium Fusarium stalk rot; ear isolates are usedgraminearum rot alone or in Fusarium Fusarium ear rot combination forverticillioides any treatment, Rhizoctonia solani Rhizoctonia crown andincluding NLS0020 root rot applied in-furrow Stenocarpella maydisDiplodia stalk and ear and NLS0109 rot applied foliar. Pythium spp.Pythium root rot Sclerospora Downy mildew; Crazy graminicola; topSclerophthora macrospora NLS0017 + Corn Pythium spp. Pythium root andstalk Seed treatment; NLS0109 rot in-furrow; foliar; or Rhizoctoniasolani Rhizoctonia crown and any combination root rot thereof, whereStenocarpella maydis Diplodia stalk and ear isolates are used rot aloneor in Sclerospora Downy mildew; Crazy combination for graminicola; topany treatment, Sclerophthora including NLS0017 macrospora appliedin-furrow Fusarium spp. Fusarium root and and NLS0109 stalk rot appliedfoliar. Colletotrichum Anthracnose leaf blight graminicola and stalk rotCercospora zeae- Gray leaf spot maydis NLS0017 + Soybean SclerotiniaWhite mold Seed treatment; NLS0109 sclerotiorum in-furrow; foliar; orCercospora sojina Frogeye leaf spot any combination Cercospora kikuchiiPurple leaf blotch and thereof, where seed stain isolates are usedSeptoria glycines Brown spot alone or in Pythium spp. Pythium root rotcombination for Fusarium spp. Fusarium seed rot, any treatment.blight/wilt, root rot, and pod and collar rot Peronospora Downy mildewmanshurica Rhizoctonia solani Rhizoctonia damping off and root rotPhytophthora sojae Root and stem rot Fusarium virguliform Sudden deathsyndrome NLS0017 + Wheat Fusarium Fusarium head blight; Seed treatment;NLS0109 graminearum seedling blight foliar; or both Pythium spp Pythiumroot rot Septoria tritici; Septoria/Stagonospora Stagonospora nodorumblotch Rhizoctonia solani Rhizoctonia root rot Fusarium spp. Fusariumroot, crown, and foot rot Magnaporthe grisea Head blast Pyrenophoratritici- Tan Spot repentis Microdochium nivale Snow mold Blumeriagraminis f. Powdery mildew sp. tritici NLS0064 + Soybean SclerotiniaWhite mold Seed treatment; NLS0109 sclerotiorum in-furrow; foliar; orCercospora sojina Frogeye leaf spot any combination Cercospora kikuchiiPurple leaf blotch and thereof, where seed stain isolates are usedSeptoria glycines Brown spot alone or in Pythium spp. Pythium root rotcombination for Fusarium spp. Fusarium seed rot, any treatment.blight/wilt, root rot, and pod and collar rot Peronospora Downy mildewmanshurica Rhizoctonia solani Rhizoctonia damping off and root rotPhytophthora sojae Root and stem rot Fusarium virguliform Sudden deathsyndrome NLS0021 + Corn Pythium spp. Pythium root and stalk Seedtreatment; NLS0109 rot in-furrow; foliar; or Rhizoctonia solaniRhizoctonia crown and any combination root rot thereof, whereStenocarpella maydis Diplodia stalk and ear isolates are used rot aloneor in Sclerospora Downy mildew; Crazy combination for graminicola; topany treatment, Sclerophthora including NLS0021 macrospora appliedin-furrow Fusarium spp. Fusarium root and and NLS0109 stalk rot appliedfoliar. Colletotrichum Anthracnose leaf blight graminicola and stalk rotCercospora zeae- Gray leaf spot maydis

In certain embodiments, an amount of a composition provided herein thatis sufficient to provide for inhibition of fungal infection in a plantor plant part can be a composition with Methylobacterium that inhibitplant pathogenic fungi at a titer of at least about 5×10⁸ colony-formingunits per milliliter, at least about 1×10⁹ colony-forming units permilliliter, at least about 1×10¹⁰ colony-forming units per milliliter,or at least about 3×10¹⁰ colony-forming units per milliliter. In certainembodiments, an amount of a composition provided herein that issufficient to provide for inhibition of fungal disease in a plant orplant part can be a composition with Methylobacterium that inhibit plantpathogenic fungi at a titer of about 5×10⁸ colony-forming units permilliliter to at least about 6×10¹⁰ colony-forming units per milliliter.In certain embodiments, an amount of a composition provided herein thatis sufficient to provide for inhibition of fungal disease in a plant orplant part can be a fermentation broth product with a Methylobacteriumthat inhibit plant pathogenic fungi titer of a solid phase of thatproduct is at least about 1×10⁷, 5×10⁷, 1×10⁸, or 5×10⁸ colony-formingunits per gram to at least about 6×10¹⁰, 1×10¹³, or 5×10¹³colony-forming units of Methylobacterium per gram of the solid phasewherein a mono-culture or co-culture of Methylobacterium that inhibitplant pathogenic fungi is adhered thereto. In certain embodiments, anamount of a composition provided herein that is sufficient to providefor inhibition of fungal disease in a plant or plant part can be acomposition with a Methylobacterium titer of at least about 1×10⁷,5×10⁷, 1×10⁸, or 5×10⁸ colony-forming units per gram to at least about6×10¹⁰, 1×10¹³, or 5×10¹³ colony-forming units of Methylobacterium pergram of particles in the composition containing the particles thatcomprise a solid substance wherein a mono-culture or co-culture ofMethylobacterium that inhibit plant pathogenic fungi is adhered thereto.In any of the aforementioned compositions, the indicated concentrationscan be fungal inhibitory concentrations.

In certain embodiments, the compositions, plants and plant parts treatedwith the compositions, and methods provided herein can comprisefungicides in addition to the Methylobacterium strains for enhanceddisease control. In certain embodiments, the fungicides are provided incompositions comprising the Methylobacterium, for example as seedtreatments, as slurries for in furrow treatment or in foliar sprays. Incertain embodiments, the fungicides are provided as separatecompositions for use in the methods provided herein. Examples offungicides that can be used in the compositions and methods providedherein for enhanced control of plant pathogenic fungi are provided inTable 4.

TABLE 4 Chemicals to Combine with Methylobacterium for Crop PathogenDisease Common Name(s) Enhanced Disease Control Wheat Fusarium Fusariumhead blight; tebuconazole, triticonazole, graminearum seedling blightmetconazole, ipconazole, prothioconazole Wheat Pythium spp Pythium rootrot metalaxyl, mefenoxam, oxathiapiprolin, ethaboxam, thiram,thiophanate-methyl Wheat Blumeria graminis f. Powdery mildewpropiconazole, triadimenfon, sp. tritici fluxapyroxad, (trifloxystrobin,fluopyram specialty crops) Wheat Septoria tritici; Septoria/Stagonosporatebuconazole, triticonazole, Stagonospora blotch metconazole,ipconazole, nodorum prothioconazole Wheat Rhizoctonia Rhizoctonia rootrot tebuconazole, triticonazole, solani metconazole, ipconazole,prothioconazole, thiram, captan, penflufen, carboxin, fluxapyroxad WheatFusarium spp. Fusarium root, crown, and tebuconazole, triticonazole,foot rot metconazole, ipconazole, prothioconazole, thiram, captan,penflufen, imazalil, carboxin, thiabendazole, fluxapyroxad WheatMagnaporthe Head blast prothioconazole, trifloxystrobin grisea WheatPyrenophora Tan Spot tebuconazole, triticonazole, tritici-repentismetconazole, ipconazole, prothioconazole Wheat Microdochium Snow molddimethomorph, boscalid nivale Corn Cercospora Gray leaf spotazoxystrobin, pyraclostrobin, zeae-maydis fluoxastrobin, andtrifloxystrobin, fenamidone Corn Colletotrichum Anthracnose (foliar andipconazole, prothioconazole, graminicola stalk rot) propiconazole,azoxystrobin, pyraclostrobin, fluoxastrobin, and trifloxystrobin CornFusarium Fusarium stalk rot; ear rot tebuconazole, triticonazole,graminearum metconazole, ipconazole, prothioconazole, thiram, captan,penflufen, imazalil, carboxin, thiabendazole, fluxapyroxad Corn FusariumFusarium ear rot tebuconazole, triticonazole, verticillioidesmetconazole, ipconazole, prothioconazole, thiram, captan, penflufen,imazalil, carboxin, thiabendazole, fluxapyroxad Corn Fusarium spp.Fusarium root and stalk rot tebuconazole, triticonazole, metconazole,ipconazole, prothioconazole, thiram, captan, penflufen, imazalil,carboxin, thiabendazole, fluxapyroxad Corn Stenocarpella Diplodia stalkand ear rot tebuconazole, triticonazole, maydis metconazole, ipconazole,prothioconazole, thiram, captan, penflufen, carboxin, fluxapyroxad CornPythium spp. Pythium root and stalk rot metalaxyl, mefenoxam,oxathiapiprolin, ethaboxam, thiram, thiophanate-methyl Corn SclerosporaDowny mildew; Crazy top propamocarb, dimethomorph, graminicola;metalaxyl, mefenoxam Sclerophthora macrospora Corn RhizoctoniaRhizoctonia crown and tebuconazole, triticonazole, solani root rotmetconazole, ipconazole, prothioconazole, thiram, captan, penflufen,carboxin, fluxapyroxad Soybean Cercospora Frogeye leaf spotazoxystrobin, pyraclostrobin, sojina fluoxastrobin, and trifloxystrobinSoybean Cercosporia Purple leaf blotch and seed azoxystrobin,pyraclostrobin, kikuchii stain fluoxastrobin, and trifloxystrobin,fenamidone Soybean Septoria Brown spot azoxystrobin, pyraclostrobin,glycines fluoxastrobin, and trifloxystrobin, fluopyram Soybean Pythiumspp. Pythium root rot metalaxyl, mefenoxam, oxathiapiprolin, ethaboxamSoybean Sclerotinia White mold tebuconazole, prothioconazole,sclerotiorum fluopyram Soybean Fusarium spp. Fusarium seed rot,tebuconazole, triticonazole, blight/wilt, root rot, and metconazole,ipconazole, pod and collar rot prothioconazole, thiram, captan,penflufen, imazalil, carboxin, thiabendazole, fluxapyroxad SoybeanFusarium Sudden death syndrome fluopyram virguliform Soybean RhizoctoniaRhizoctonia damping off tebuconazole, triticonazole, solani and root rotmetconazole, ipconazole, prothioconazole, thiram, captan, penflufen,carboxin, fluxapyroxad Soybean Phytophthora Root and stem rot metalaxyl,mefenoxam, sojae oxathiapiprolin

When chemicals are used in combination with Methylobacterium,application rates of the chemicals may be reduced compared to standardcommercial application rates for a given plant species, while providingthe same or increased level of protection to the plant from plantpathogenic fungal pathogens. In certain embodiments, Methylobacteriumstrains provided here provide enhanced activity in combination withfungicides as the result of activity against particular fungal strainsfor which the chemical fungicides provide no or incomplete control. Incertain embodiments, the Methylobacterium is NLS0109 or NL0089, thefungus is Cercospora, the crop is corn, the disease is grey leaf spot,and the chemical is a strobilurin, such as pyraclostrobin, or an azole,such as tebuconazole. In certain embodiments, the fungicide applicationrate or numbers of applications of the fungicide can be reduced as theresult of antifungal activity of the applied Methylobacterium. Incertain embodiments, the Methylobacterium is NLS0109, the fungus isPythium, the crop is soybean or corn, the disease is Pythium root rot,and the chemical is metalaxyl, mefenoxam, oxathiapiprolin or ethaboxam.In certain embodiments, the fungicide application rate or numbers ofapplications of the fungicide can be reduced as the result of antifungalactivity of the applied Methylobacterium.

EXAMPLES

The following examples are included to demonstrate various embodiments.It will be appreciated by those of skill in the art that the techniquesdisclosed in the following examples represent techniques discovered bythe Applicants to function well. However, those of skill in the artshould, in light of the instant disclosure, appreciate that many changescan be made in the specific embodiments that are disclosed, while stillobtaining like or similar results, without departing from the scope ofthe disclosure.

Example 1 Corn and Soybean Field Trials Summer of 2015

In the summer of 2015, field trials to evaluate disease suppression incorn and soybeans by PPFMs were performed at two independent locations:Bethel, Mo. and Troy, Ohio. Both trial locations were managed bycontract research organizations. NewLeaf Symbiotics personnel visitedeach site at least twice to ensure proper trial implementation. The samestrains and application rates were tested at both locations. The trialswere arranged as a split-plot within an RCBD (randomized complete blockdesign) with six replications at the Bethel site and four replicationsat the Troy site. Treatments for corn are described in Table 5 andtreatments for soybean are described in Table 6. In-furrow treatmentswere applied at a rate of 1,250 mL 10× PPFM concentrate per acre andfoliar treatments were applied at a rate of 5,000 mL 10× PPFMconcentrate per acre. The split-plot design allowed for the evaluationof in-furrow treatment, foliar treatment, response to sequential PPFMtreatments, and interactions between different PPFMs. Data from theTroy, Ohio soybean trial were not analyzed.

TABLE 5 2015 Pathology Corn Field Trial Treatments Treatment NumberWhole-plot treatment Sub-plot treatment 1 Mock Mock 2 NLS0020 Mock 3Mock NLS0020 4 NLS0020 NLS0020 5 Mock NLS0109 6 NLS0020 NLS0109 7 MockNLS0066 8 NLS0020 NLS0066

TABLE 6 2015 Pathology Soybean Field Trial Treatments Treatment NumberWhole-plot treatment Sub-plot treatment 1 Mock Mock 2 NLS0089 Mock 3Mock NLS0020 4 NLS0089 NLS0020 5 Mock NLS0109 6 NLS0089 NLS0109 7 MockNLS0066 8 NLS0089 NLS0066

At each site, conventional row spacing was used and standard agronomicpractices were followed. Corn and soy hybrids with similar genetics butsuitable for the specific trial locations were supplied for each site.Sub-plot sizes were no less than four 20′ rows. A five-foot border wasleft between sub-plot to mitigate neighbor effects. Additionally,observations were taken from only the center two rows of each plot.Whole-plots consisted of the four sub-plots plus five foot bordersbetween plots. Trial locations were selected in areas with naturaldisease pressure and no artificial inoculations were made. As a result,the same diseases were not evaluated at each location. Diseases rated incorn were anthracnose (Colletotrichum graminicola), grey leaf spot(Cercospora zeae-maydis), and common rust (Puccinia sorghi). Diseasesrated in soybean were brown spot (Septoria glycines) and various foliardiseases, particularly frogeye leafspot (Cercospora sojina). For eachdisease present, incidence and/or severity ratings were collected andanalyzed to determine treatment effects.

Disease ratings and statistical analysis results are reported in Tables7-9. Due to the different disease ratings and replication number at eachsite, data from the two trial locations were analyzed separately. Dataanalyses were performed using SAS JMP software v11.2 (SAS Institute,Cary, N.C.). Data were analyzed according to JMP guidelines forsplit-plot analysis within the ‘Fit Model’ function, which uses the REMLtechnique for mixed models. Student's T and Tukey's HSD post hoc testswere applied to determine differences between treatment groups (α=0.05).Contrasts were used to make comparisons between specific groups ofinterest.

TABLE 7 Soybean Foliar Disease-Bethel, Missouri Brown Brown spot spotLeaf spot Leaf spot Whole-plot Sub-plot severity severity severityseverity (in-furrow) (foliar) early late early late Treatment Treatment(%) (%) (%) (%) Mock Mock 3.00 8.33 1.67 5.50 Mock NLS0020 1.33^(T)4.17^(T,H) 0.17^(T,H) 2.00^(T,H) Mock NLS0066 1.67^(T) 4.83^(T) 0.67^(T)4.17 Mock NLS0109 1.67^(T) 5.67^(T) 0.50^(T,H) 4.00^(T) NLS0089 Mock1.17^(T) 5.00^(T) 0.17^(T,H) 2.50^(T,H) NLS0089 NLS0020 1.17^(T)5.17^(T) 0.33^(T,H) 2.50^(T,H) NLS0089 NLS0066 1.67 5.17^(T) 0.33^(T,H)4.33 NLS0089 NLS0109 2.00 5.00^(T) 0.33^(T,H) 3.00^(T) ^(T)Treatmentsignificantly different from control (Mock, Mock) by Student's T-test (α= 0.05) ^(H)Treatment significantly different from control (Mock, Mock)by Tukey's HSD (α = 0.05)

TABLE 8 Corn Foliar Disease-Bethel, Missouri Anthrac- Gray leaf Grayleaf Common Whole-plot Sub-plot nose spot spot rust (in-furrow) (foliar)severity severity severity severity Treatment Treatment (%) early (%)late (%) (%) Mock Mock 21.17 3.17 13.00 12.17 Mock NLS0020 18.832.00^(T,H) 11.33 11.67 Mock NLS0066 9.50^(T,H) 1.83^(T,H) 10.83^(T)10.50 Mock NLS0109 9.67^(T,H) 2.50^(T) 11.00^(T) 10.00^(T) NLS0020 Mock18.67 2.17^(T) 11.83 10.67 NLS0020 NLS0020 17.83 2.00^(T) 11.50 11.33NLS0020 NLS0066 9.17^(T) 1.00^(T,H) 9.50^(T,H) 9.67^(T) NLS0020 NLS01099.17^(T) 1.17^(T,H) 9.83^(T,H) 9.17^(T,H) ^(T)Treatment significantlydifferent from control by Student's T-test (α = 0.05) ^(H)Treatmentsignificantly different from control by Tukey's HSD (α = 0.05)

TABLE 9 Corn Foliar Disease-Troy, Ohio Gray leaf Tip Tip StalkWhole-plot Sub-plot spot dieback dieback rot (in-furrow) (foliar)severity severity incidence severity Treatment Treatment (%) (%) (%) (%)Mock Mock 67.50 11.00¹ 0.17¹ 1.55 Mock NLS0020 67.50 8.25 0.14 1.60 MockNLS0066 62.50 12.50 0.18 1.45 Mock NLS0109 60.00 11.75 0.19 1.40 NLS0020Mock 65.00 7.50* 0.12* 1.85 NLS0020 NLS0020 67.50 10.00 0.16 1.55NLS0020 NLS0066 60.00 9.00 0.14 1.45 NLS0020 NLS0109 62.50 9.25 0.141.85 ¹The average across all mock in-furrow treatments was significantlydifferent from the average across all NLS0020 in-furrow treatments bycontrast (α = 0.05) *Treatment significantly different from control(Mock, Mock) by contrast (α = 0.10)

All treatments applied to soybeans demonstrated disease suppressionagainst both brown spot (Septoria glycines) and other foliar leaf spotdiseases. Foliar application of NLS0020 without in-furrow treatmentresulted in the lowest rating for all diseases and was the mosteffective treatment for suppression of disease relative to the control.Foliar application of NLS0020 following NLS0089 in-furrow treatment alsodemonstrated disease suppression across all treatments. Foliar treatmentof NLS0109 significantly reduced all diseases and provided greatersuppression when applied after in-furrow treatment with NLS0089 for allapplications except the early brown spot rating. In-furrow treatmentwith NLS0089 alone significantly reduced all diseases and had aparticularly strong effect against foliar leaf spot diseases.

In corn at the Bethel, Missouri site, foliar applications of NLS0066 andNLS0109 significantly suppressed all diseases, with the exception ofNLS0066 against common rust. In-furrow application of NLS0020 improvedthe disease suppression provided by NLS0066 and NLS0109 foliarapplications in all examples. This demonstrates enhanced efficacythrough multiple applications of these specific PPFM strains.Application of NLS0020 alone, including foliar, in-furrow, and in-furrowfollowed by foliar, resulted in reduction of early Gray leaf spotseverity, but had no significant impact on the other diseases tested.

At the Troy, Ohio location, the NLS0066 and NLS0109 foliar applicationsprovided the lowest gray leaf spot severity ratings. This was inagreement with the treatment effects observed in Missouri. In-furrowapplication of NLS0020 alone suppressed both the severity and incidenceof tip dieback, which can be indicative of an effect on disease andabiotic stressors. Additionally, all applications of in-furrow NLS0020combined suppressed tip dieback metrics relative to all mock in-furrowtreatments combined, indicating an overall positive effect of NLS0020in-furrow treatment.

Example 2 Suppression of White Mold by Methylobacterium

Sclerotinia sclerotiorum is a polyphagous ascomycete fungus, with a hostrange that encompasses thousands of dicot plants. White mold on soybeanand other leguminous crops is of particular agronomic importance. Undercool, moist environmental conditions, white mold causes prematuresenescence and drastically reduced yields. There is no availablecomplete genetic resistance to white mold and partial resistance is onlymarginally effective. Further, fungicide applications specifically forwhite mold are only applied in years when disease is highly likely andmust be applied within a narrow window in order to provide effectiveprotection. Application of beneficial bacteria that colonize the plantand provide enhanced disease resistance could mitigate effects of whitemold and increase the efficacy of genetic and chemical diseasemanagement tactics.

Frozen PPFM stock solutions at a 10× concentration of approximately1×10⁸ CFU/mL were thawed to room temperature directly prior to foliartreatment. PPFM cells were washed by centrifuging to pellet the cells,pouring off the resulting supernatant, re-suspending the cells in one mLsterile distilled water, centrifuging and pouring off the supernatant asecond time, then re-suspending a second time in 1 mL of steriledistilled water. Re-suspended, washed cells were then diluted to a 1×concentration. Within two hours of PPFM preparation, an airbrushcalibrated to 20 psi was used to apply 10 mL of 1× PPFM solution evenlyacross each flat of treatment plants. PPFMs were applied to three-weekold plants and inoculations were performed the following week whenplants were approximately one-month old.

Seeds were planted into either potting media, field soil, or a 50/50 mixof potting media and field soil, depending on the specific experiment.In all experiments, flats holding 18 pots each were used and the potscontaining individual treatments were organized into randomized completeblocks either at planting or just prior to inoculation. Immediatelyafter planting, pots were moved to a greenhouse (75-80° F.; RH 40-90%;16 h day-length) and grown there for one month. Plants were watereddaily and received supplemental fertilizer two times per week.

One-month old plants were inoculated with 5-7 day-old cultures ofSclerotinia sclerotiorum grown on PDA in the dark. A modified version ofthe cut petiole inoculation technique was used (Hoffman et al. 2002.Plant Dis. 86:971-980). Briefly, the petiole of the third trifoliate wascut with scissors approximately one inch from the stem. The broad end ofa 1000 uL pipet tip was used to excise an agar disk from the outer edgeof an S. sclerotiorum culture. The tip was then placed over the cutpetiole such that the broad end of the pipet tip was in contact with thestem and petiole base and the cut end of the petiole was in contact withthe mycelium side of the agar plug. A small piece of parafilm waswrapped around the tip and stem to prevent the tip from falling off.Inoculated plants were incubated in the greenhouse for 7-10 days toallow for disease development prior to rating.

Lesion length and wilt severity were collected as disease metrics.Length of brown or bleached lesions was measured using a ruler. Wiltseverity was rated on a 0-5 scale with 0 indicating a completely healthplant and 5 indicating a dead plant.

A total of 12 different PPFM strains in five individual experiments,each replicated three times, were tested for suppression of white moldfollowing foliar PPFM application (Table 10). Of these strains, onlyNLS0020 and NLS0109, both of which were in the first two experiments,had a significant effect on either lesion length or severity of wiltcaused by pathogen infection (Table 11). In the first experiment, whichhad a lower sample size, no significant effects were detected but bothNLS0020 and NLS0109 decreased white mold lesion length and wilt severityby at least 20% relative to the mock-treated control. In the secondexperiment, NLS0020 decreased lesion length by approximately 30%(P=0.04) but did not have a significant effect on wilt severity (10%reduction; P=0.76). NLS0109 decreased lesion length by approximately 40%(P<0.01) and wilt by approximately 40% (P=0.06). For analysis, aDunnett's test was conducted in the ‘Fit Y by X’ function of SAS JMPv12.0 statistical analysis software. The mock-foliar treatmentapplication group, which received a bacterial growth medium only foliarapplication, was designated as the control group.

TABLE 10 Lesion Length (LL) and Wilt Severity of White Mold FollowingFoliar Application of PPFM Strains Strain LL ± SE LL RTC (%) Wilt ± SEWilt RTC (%) Experiment 1 Control 100.42 ± 9.55  100.00 4.46 ± 0.26100.00 NLS0020  74.50 ± 10.65 74.19 3.33 ± 0.39 74.67 NLS0071  84.79 ±12.66 84.44 3.54 ± 0.41 79.37 NLS0109  79.17 ± 09.90 78.84 3.58 ± 0.4180.26 Experiment 2 Control 106.04 ± 11.18 100.00 4.13 ± 0.34 100.00NLS0020 74.17 ± 8.67 69.94 3.71 ± 0.35 89.90 NLS0071  94.38 ± 10.7789.00 4.12 ± 0.32 101.01 NLS0109  64.38 ± 10.81 60.71 2.92 ± 0.45 70.71Experiment 3 Control 45.00 ± 6.12 100.00 2.19 ± 0.33 100.00 NLS001755.33 ± 7.12 123.00 2.83 ± 0.32 129.00 NLS0037 50.63 ± 7.02 112.50 2.19± 0.38 100.00 NLS0089 45.16 ± 6.52 100.35 1.94 ± 0.33 88.57 Experiment 4Control 63.75 ± 8.16 100.00 3.22 ± 0.33 100.00 NLS0046 54.22 ± 9.3385.05 2.34 ± 0.33 72.82 NLS0064 67.81 ± 9.07 106.37 2.88 ± 0.34 89.32NLS0066 59.38 ± 9.03 93.13 2.56 ± 0.35 79.61 Experiment 5 Control 79.38± 7.52 100.00 3.72 ± 0.30 100.00 IP0021 97.26 ± 7.35 122.53 4.39 ± 0.21117.97 IP0121 95.78 ± 6.77 120.67 4.16 ± 0.17 111.76 IP0237 84.38 ± 6.58106.30 4.16 ± 0.24 111.76

TABLE 11 Analysis of White Mold Control by PPFM Strains LL Wilt Strain(% control) LL p-value (% control) Wilt p-value NLS0071 11.00 0.6975−1.01 0.9996 NLS0020 30.06 0.0429 10.10 0.7625 NLS0109 39.29 0.005329.29 0.0589

Foliar applications of PPFM strain NLS0109 reduced symptoms of bothlesion length and wilt on soybean caused by the white mold pathogenSclerotinia sclerotiorum. Applications of this strain could be used as astandalone treatment for suppression of white mold. Further, NLS0109could be mixed with chemical or biological disease management tools aspart of an integrated disease management approach. Such combinationscould be leveraged to increase and/or prolong product efficacy and aspart of a resistance management program.

Example 3 Suppression of Gray Leaf Spot by Methylobacterium spp. inCombination and Comparison With Conventional Fungicides

Gray leaf spot, caused by the ascomycete fungal pathogen Cercosporazeae-maydis, is a serious disease of field corn. While varying levels ofresistance to this disease are present in commercial germplasm, nocomplete resistance is available. Gray leaf spot is a perennial problemand is particularly severe during summers with high relative humidity.The summer of 2016 was favorable for gray leaf spot, with diseasepressure starting in June and persisting throughout the growing season.

In this trial, a greenhouse study was conducted to determine theefficacy of PPFM strains when applied alone, in combination with eachother, or in combination with low label rates of the foliar fungicideHeadline™ (active ingredient—pyraclostrobin; BASF Corporation, ResearchTriangle, N.C., USA). These treatments were compared against Headlineapplication alone, as well as other agriculturally relevant fungicidetreatment and treatment combinations containing pyraclostrobin as anactive ingredient (Table 12).

TABLE 12 PPFM/Industry Standard Greenhouse Trial Treatments No.Treatment Rate Timing 1 Untreated control 0 fl oz/A N/A 2Headline ™-Foliar 6 fl oz/A VT 3 Fortix ™ ¹-Foliar 4 fl oz/A VT 4Xanthion ™ ²-In-furrow 0.6 fl oz/A Component A Planting 3.0 fl oz/AComponent B 5 Xanthion ™ In-furrow 0.6 fl oz/A Component A PlantingHeadline ™-Foliar 3.0 fl oz/A Component B 6 fl oz/A VT 6NLS0020-In-furrow 1,250 mL/A Planting 7 NLS0020-In-furrow 1,250 mL/APlanting Headline ™-Foliar 6 fl oz/A VT 8 NLS0109-Foliar 5,000 mL/A VT 9NLS0109-Foliar 5,000 mL/A VT Headline ™-Foliar 6 fl oz/A VT 9NLS0020-In-furrow 1,250 mL/A Planting NLS0109-Foliar 5,000 mL/A VT 10NLS0020-In-furrow 1,250 mL/A Planting NLS0109-Foliar 5,000 mL/A VTHeadline ™-Foliar 6 fl oz/A VT ¹ Fortix ™ (Arysta Life Sciences, Cary,North Carolina, USA) ² Xanthion ™ (BASF Corporation, Research Triangle,NC, USA)

The trial was conducted in Spring-Summer 2016 with corn planted in earlyApril and inoculated two-weeks prior to anthesis. Inoculum was appliedby placing pathogen-infected corn debris around the base of each plantand then increasing relative humidity within the greenhouse to 60-90%during the day and greater than 90% at night, with the temperatureranging from 70-82° F. To promote infection, a gray leaf spotsusceptible corn hybrid (Sun Prairie 617RR) was used for the trial.Disease ratings were taken at 14, 21, and 28 days post-inoculation and afinal ear weight was collected for each plant.

The trial was arranged as an RCBD with 10 replications. In-furrowtreatments were applied at planting using a hand-held sprayer andapplication volumes were scaled down to the total pot area. Fungicideswere mixed and applied in accordance with label instructions. PPFMs weresupplied as frozen concentrate that was thawed just prior toapplication. Extra seeds were planted and treated for each treatmentgroup to ensure a total of ten replicates were available forinoculation. Two weeks after germination, corn seedlings weretransferred from seedlings trays to 2.5 gallon pots, in which they grownfor the remainder of the experiment. Foliar applications were performedat the VT growth-stage using a backpack sprayer. To prevent drift withinthe greenhouse, plants in each treatment group were brought outside andthe treatment was applied to the entire group. Plants were then returnedto the greenhouse and randomized into the ten experimental replicates.

Disease data were analyzed in JMP version 12.0 (SAS Institute, Cary,N.C.). Disease incidence and severity values collected at each samplingdata were used to calculate disease index [(incidence*severity)/100] anda cumulative area under the disease progress curve (AUDPC) wascalculated the three disease index values for each treatment. Data wereanalyzed using the ‘Fit Model’ function in JMP with ‘Rep’ as a randomeffect and ‘Treatment’ as a fixed effect. A post-hoc Tukey test at analpha level of 0.05 was applied to perform means separation betweentreatments (Table 13).

PPFM treatments alone did not perform differently from the UTC or fromin-furrow treatment with Xanthion™. Xanthion™ is a combination productof pyraclostrobin and the Bacillus subtilis microbial inoculant strainMBI600; thus, PPFM strains alone performed similarly to a currentlyavailable commercial product that combines a chemical fungicide with amicrobial agent. The combination of NL0020-IF/NL0109-F reduced diseaserelative to the UTC and disease levels under this treatment were notsignificantly different from those of either foliar-appliedpyraclostrobin-based fungicide (Headline™ and Fortix™).NL0020-IF/Headline-F™ suppressed disease to levels similar to those ofXanthion-IF™/Headline-F™, Headline-F™, and Fortix-F™. As theXanthion™/Headline™ combination requires two applications ofpyraclostrobin, similar performance by NLS0020/Headline indicates thatan application of this active ingredient could be dropped in favor of aPPFM application without any adverse effects on disease severity.

Of all treatments, NLS0109-F/Headline-F™ andNLS0020-IF/NLS0109-F+Headline-F™ had the greatest effects on diseasesuppression relative the UTC. In particular, these treatmentsnumerically reduced disease to levels greater than Headline™ alone atthe 21- and 14-day rating intervals (FIG. 4). This indicates thepotential for PPFM applications, particularly NLS0109 foliarapplications made in combination with Headline™, to extend the window ofprotection provided by conventional fungicides used for gray leaf spotmitigation.

In addition to disease suppression, PPFM application had a strongpositive effect on ear weight (Table 12). The average weight for alltreatments with PPFMs was 132.67 grams/ear, which is nearly ten gramshigher than the 123.00 grams/ear average of the untreated control andfungicide-only treatments. Fortix™ alone was the only fungicidetreatment to perform similarly to PPFM treatments and NLS0020 in-furrowapplication was particularly conducive to increased yield weight,garnering three of the four highest weights obtained.

This experiment demonstrates the potential for use of PPFMs incombination with conventional fungicides. PPFMs can be used to increaseproduct efficacy by prolonging the window of effective protectionoffered by currently available chemistries and by offering an alternateproduct for use in resistance management. Additionally, PPFMs had apositive effect on ear weight, indicating a potential to boost yieldswhen used alone or in combination with conventional fungicides.

TABLE 13 Gray Leaf Spot Greenhouse Evaluation Results % Dif- % Dif-Yield ference ference LSMeans (grams/ from from Comparison Treatmentear) UTC AUDPC UTC (Tukey) UTC 120 0.00 39.55 0.00 A Headline-F ™ 1210.83 5.57 −85.92 BC Fortix-F ™ 135 12.50 6.3 −84.07 BC NLS0109-F 1275.83 28.49 −27.96 A NLS0109-F + 129 7.50 3.15 −92.04 C Headline-F ™Xanthion-IF ™ 116 −3.33 38.64 −2.30 A Xanthion-IF ™/ 123 2.50 5.67−85.66 BC Headline-F ™ NLS0020-IF 135 12.50 35.39 −10.52 A NLS0020-IF/138 15.00 7.72 −80.48 BC Headline-F ™ NLS0020-IF/ 131 9.17 16.44 −58.43B NLS0109-F NLS0020-IF/ 136 13.33 3.12 −92.11 C NLS0109-F + Headline-F ™

Example 4 Grey Leaf Spot Field Trials Summer 2016

A second year of corn pathology field trials was conducted in 2016.Trials were placed in sites with naturally high levels of gray leaf spotinoculum and a susceptible corn hybrid (Sun Prairie 617RR) was used tofacilitate gray leaf spot infection. Two trial locations were included:Dana, Iowa and Bethel, Mo. At each site, the trial was conducted as arandomized complete block with six replicates. Standard local agronomicpractices for fertilizer application, tillage, row spacing, population,and pest management were used. Standard weed and insect managementpractices were employed and seed was supplied to cooperators pre-treatedwith a standard fungicide/insecticide seed treatment package (Acceleron™2016 Corn, Monsanto, St. Louis, Mo., USA). Foliar fungicide applicationswere only made if specified in the trial protocol. Sixteen treatmentswere included in the trials, including PPFMs applied alone, PPFMsapplied in combination, PPFMs applied alongside chemical fungicides, andfungicide alone standards (Table 14). PPFM compositions were diluted toa concentration of approximately 10⁹ CFU/ml and applied at the ratesshown in Table 14. Assessments collected included early season stand andvigor, mid-season gray leaf spot incidence and severity, flowering date,stalk diameter, lodging, final stand at harvest, yield, and moisturecontent. Incidence and severity data from each rating date were combinedto provide a disease index value [(incidence*severity)/100]. Data fromeach site were analyzed separately used JMP v12.0 (SAS Institute, Cary,N.C.). Treatment effects on response variables of interest weredetermined using the ‘Fit Model’ function with ‘Treatment’ specified asa fixed effect and ‘Rep’ as a random effect. Post-hoc means separationwere performed using the ISMeans Differences Student's T′ option within‘Fit Model’ and means are reported as LSMeans from the correspondingmodel. At the Bethel, Missouri site the Untreated check was the lowestyielding treatment and maintained the highest levels of gray leaf spotdisease incidence and severity throughout the growing season (Table 15).Treatments with NLS0109 provided a greater than 10 bu/A yield increaseover the Untreated check. Disease index values for both treatments withNLS0109 were significantly lower (P<0.05) than the Untreated check onthe first two rating dates (August 12 and August 21) but on the finalrating date (August 28) only the treatment combination of NLS0020in-furrow followed by an NLS0109 foliar treatment suppressed diseaserelative to the Untreated check (Table 15).

TABLE 14 2016 Gray Leaf Spot Field Trial Treatments Trt No Treatment(s)Application Timing Rate 1 Untreated Control N/A N/A 2 Headline ™ VT¹ 6.0fl oz/A 3 Fortix ™ VT 4/0 fl oz/A 4 Xanthion ™ In-furrow @ planting 0.6fl oz/A Component A 3.0 fl oz/A Component B Headline ™ VT 6 fl oz/A 5NLS0020 In-furrow @ planting 1.25 L/A 6 NLS0020 In-furrow @ planting1.25 L/A NLS0066 VT 5.0 L/A 7 NLS0020 In-furrow @ planting 1.25 L/ANLS0089 VT 5.0 L/A 8 NLS0020 In-furrow @ planting 1.25 L/A NLS0109 VT5.0 L/A 9 NLS0020 In-furrow @ planting 1.25 L/A Headline ™ VT 6 fl oz/A10 NLS0020 In-furrow @ planting 1.25 L/A Fortix ™ VT 4.0 fl oz/A 11NLS0021 In-furrow @ planting 1.25 L/A 12 NLS0021 In-furrow @ planting1.25 L/A NLS0066 VT 5.0 L/A 13 NLS0021 In-furrow @ planting 1.25 L/ANLS0089 VT 5.0 L/A 14 NLS0021 In-furrow @ planting 1.25 L/A NLS0109 VT5.0 L/A 15 NLS0021 In-furrow @ planting 1.25 L/A Headline ™ VT 6.0 floz/A 16 NLS0021 In-furrow @ planting 1.25 L/A Fortix VT 4.0 fl oz/A ¹VTstage of development: last branch of tassel completely visible but silksnot visible.

TABLE 15 Summer 2016 Gray Leaf Spot Field Trial Results-Bethel, MissouriTreatment Yield Index_12 Aug. Index_21 Aug. Index_28 Aug. UntreatedCheck 141.77¹ C² 7.25 A 20.83 A 24.83 A NLS0020 149.93 BC 4.85 B 15.27 B21.33 AB NLS0020/NLS0066 161.20 AB 2.83 C 12.28 BCD 20.07 BCNLS0020/NLS0089 159.88 AB 2.08 CD 12.37 BCD 18.53 BC NLS0020/NLS0109155.15 ABC 2.65 CD 10.97 CD 19.07 BC NLS0020/Fortix ™ 160.32 AB 0.38 F1.78 E 8.55 D NLS0020/Headline ™ 161.62 AB 0.83 EF 2.17 E 5.88 D NLS0021157.32 AB 4.00 B 13.15 BC 19.88 BC NLS0021/NLS0066 159.47 AB 1.57 DE8.87 D 16.3 C NLS0021/NLS0089 152.57 BC 2.18 CD 11.3 BCD 20.58 ABCNLS0021/NLS0109 153.40 ABC 2.45 CD 11.62 BCD 20.88 AB NLS0021/Fortix ™164.22 AB 0.25 F 1.03 E 6.28 D NLS0021/Headline ™ 167.23 A 0.23 F 1.57 E4.83 D Fortix ™ 161.62 AB 0.4 F 1.5 E 7.23 D Headline ™ 163.78 AB 0.42 F1.2 E 5.57 D Xanthion ™ 167.27 A 0.17 F 0.9 E 5.52 D ¹Values reportedare means from six replicated plots per treatment ²Means followed by thesame capital letter are not significantly different from one another byLSMeans Differences Student's T post-hoc means comparison test (alpha =0.05)

Example 5 Detection of Methylobacterium Isolates on Target Crops

Assays are disclosed for detection of specific Methylobacterium strainsand closely related derivatives on target crops.

A qPCR Locked Nucleic Acid (LNA) based assay for NLS109 is developed asfollows. NLS109 genomic DNA sequence is compared by BLAST analysis ofapproximately 300 bp fragments using a sliding window of from 1-25nucleotides to whole genome sequences of over 1000 public andproprietary Methylobacterium isolates. Genomic DNA fragments wereidentified that had weak BLAST alignments, indicative of approximately60-95% identity over the entire fragment, to corresponding fragmentsfrom NLS0109. Target fragments from the NLS0109 genome corresponding tothe identified weak alignments regions that were selected for assaydevelopment are provided as SEQ ID NOS:9-11

TABLE 16 Target Fragment Sequences of NLS0109 SEQ ID Fragment NOSequence ref1_135566 9 ACGGTCACCCCACGGACTGGGCGAGTACCTCACCGGTGTTCTATCATAACGCCGAGTTAGTTTTCGACCGTCCCTTATGCGATGTACCACCGGTGTCGGCAGCCGATTTCGTCCCACCGGGAGCTGGCGTTCCGGTTCAGACCACCATCATCGGTCACGATGTCTGGATTGGACACGGGGCCTTCATCTCCCCCGGCGTGACTATAGGAAACGGCGCGATCGTCGGGGCCCAGGCGGTCGTCACAAGAGATGTCCCACCCTATGCGGTAGTTGCTGGCGTCCCCGC GACCGTACGACGAT ref1_135772 10CCAATAAAAGCGTTGGCCGCCTGGGCAACCCGATCCGAGCCTAAGACTCAAAGCGCAAGCGAACACTTGGTAGAGACAGCCCGCCGACTACGGCGTTCCAGCACTCTCCGGCTTTGATCGGATAGGCATTGGTCAAGGTGCCGGTGGTGATGACCTCGCCCGCCGCAAGCGGCGAATTACTCGGATCAGCGGCCAGCACCTCGACCAAGTGTCGGAGCGCGACCAAAGGGCCACGTTCGAGGACGTTTGAGGCGCGACCAGTCTCGATAGTCTCATCGTCGC GGCGAAGCTGCACCTCGA ref1_16947011 CGATGGCACCGACCTGCCATGCCTCTGCCGTCCGCGCCAGAATGGTAAAGAGGACGAAGGGGGTAAGGATCGTCGCTGCAGTGTTGAGCAGCGACCAGAGAAGGGGGCCGAACATCGGCATCAAACCTCGATTGCCACTCGGACGCGAAGCGCGTCTTGAAGGAGGGATGGAAGCGAAACGGCCGCAGAGTAACCGCCGACGAAAGATTGCACCCCTCATCGAGCAGGATCGGAGGTGAAGGCAAGCGTGGGTTATTGGTAAGTGCAAAAAATATAATGG TAGCGTCAGATCTAGCGTTC

Regions in SEQ ID NOS:9-11 where corresponding regions in otherMethylobacterium strains were identified as having one or morenucleotide mismatches from the NLS109 sequence were selected, and qPCRprimers designed using Primer3 software (Untergasser et al., 2012,Koressaar et al., 2007) to flank the mismatch regions, have a meltingtemperature (Tm) in the range of 53-58 degrees, and to generate a PCRDNA fragment of approximately 100 bp. The probe sequence was designedwith a 5′ FAM reporter dye, a 3′ Iowa Black FQ quencher, and containsone to six LNA bases (Integrated DNA Technologies, Coralville, Iowa). Atleast 1 of the LNA bases is in the position of a mismatch, while theother LNA bases are used to raise the Tm. The Tm of the probe sequenceis targeted to be 10 degrees above the Tm of the primers.

Primer and probe sequences for detection of specific detection ofNLS0109 are provided as SEQ ID NOS: 12-20 in Table 17. Each of theprobes contains a 5′ FAM reporter dye and a 3′ Iowa Black FQ quencher.

TABLE 17 Primer and Probe Sequences for Specific Detection of NLS0109SEQ ID Primer/Probe NO Sequence* NLS0109_ref1_135566_forward 12CCTCACCGGTGTTCTATCATAAC NLS0109_ref1_135566_reverse 13CCGATGATGGTGGTCTGAAC NLS0109_ref1_135566_probe 14 CGTCCC TTATGCGATGTACCA NLS0109_ref1_135772_forward 15 GATCCGAGCCTAAGACTCAAAGNLS0109_ref1_135772_reverse 16 GACCAATGCCTATCCGATCAANLS0109_ref1_135772_probe 17 AACACTTGG TAG AGACAGCCNLS0109_ref1_169470_forward 18 AAGGAGGGATGGAAGCGAAACNLS0109_ref1_169470_reverse 19 ATAACCCACGCTTGCCTTCNLS0109_ref1_169470_probe 20 CGC AG AGTAACCGCCGACGAA *Bold andunderlined letters represent the position of an LNA base

Use of Primer/Probe Sets on Isolated DNA to Detect NLS0109 andDistinguish From Related Methylobacterium Isolates

A qPCR reaction is conducted in 20 ul and contains 10 ul of 2×KiCqStart™ Probe qPCR ReadyMix™, Low ROX™ from Sigma (Cat#KCQS05-1250RXN), 1 ul of 20× primer-probe mix (final concentration ofprimers is 0.5 uM each and final concentration of probe is 0.25 uM), and9 ul of DNA template/water. Approximately 30-40 ng of DNA template isused per reaction. The reaction is conducted in a Stratagene Mx3005PqPCR machine with the following program: 95° C. for 3 min, then 40cycles of 95° C. for 15 sec and 60° C. for 1 min. The MxPro software onthe machine calculates a threshold and Ct value for each sample. Eachsample was run in triplicate on the same qPCR plate. A positive resultis indicated where the delta Ct between positive and negative controlsis at least 5.

Use of the three primer/probe sets to distinguish NLS0109 from closelyrelated isolates by analysis of isolated DNA is shown in Table 18 below.The similarity score shown for the related isolates takes into accountboth the average nucleotide identity and the alignment fraction betweenthe isolates and NLS0109. One of the tested strains, NLS0730, was usedas an additional positive control. It was isolated from a culture ofNLS0109, was confirmed by full genome sequencing as identical toNLS0109, and scored positive in all three reactions. The similarityscore of greater than 1.000 for this strain is likely the result of aslightly different assembly of the genome for this isolate compared toNLS0109. The delta Ct of approximately 15 or more between the NLS0109and NLS0730 isolates and the water only control is consistent with thesequence confirmation of the identity of these isolates. Analysis ofother isolates that are less closely related to NLS0109 results in deltaCt values similar to those for the water only control.

TABLE 18 Similarity score to Average Ct Value NLS# NLS0109 Ref1_135566Ref1_135772 Ref1_169470 NLS0730 1.005 21.08 21.31 20.35 NLS0109 1 21.9722.62 22.08 NLS0731 0.181 No Ct 37.85 >37.91 NLS0644 0.87 >36.8  >38.31No Ct NLS0700 0.88 >38.36  >38.36 >38.44 NLS0710 0.894 NoCt >37.47 >38.13 NLS0834 0.852 37.81 No Ct 37.97 NLS0939 0.862 37.9438.37 >38.35 NLS0947 0.807 38.44 No Ct No Ct NLS1015 0.894 38.77 NoCt >37.91 NLS1217 0.872 37.64 37.20 37.96 H2O only >38.14  >35.92 >37.12

Use of Primer/Probes for Detection of NLS109 on Treated Plant Materials.

Detection of NLS0109 on seed washes from treated soybean seeds.

NLS0109 can be detected and distinguished from other Methylobacteriumisolates on treated soybean seeds as follows. Soybean seeds were treatedwith Methylobacterium isolates from 10× frozen glycerol stock to obtaina final concentration of 10⁶ CFU/seed. Becker Underwood Flo Rite 1706polymer is used to improve adhesion. An uninoculated control containingpolymer and water is used. DNA is isolated from the seeds as follows.Approximately 25 ml of treated seeds are submerged for 5 minutes in 20ml 0.9% sterile saline. Tubes are vortexed for 15 minutes, then the seedwash is removed to a new tube. An additional 10 ml 0.9% sterile salineis added to the same seeds, vortexed briefly, and combined with theprevious seed wash. The seed wash liquid is centrifuged. The loosepellet is saved and transferred to smaller tubes, while the supernatantis discarded. The sample is centrifuged again, and the final sampleobtained as an approximately 100 ul loose pellet. The 100 ul pellet isused as the input for DNA extraction using MOBio UltraClean MicrobialDNA Extraction kit Cat#12224-250. As shown in Table 19, NLS0109 andNLS0730, are detected in seed washes from treated soybean seeds usingall 3 primer probe sets, as demonstrated by delta Ct of greater than 10as compared to Ct values of negative controls.

TABLE 19 Similarity score to Average Ct Value Treatment NLS0109Ref1_135566 Ref1_135772 Ref1_169470 NLS0109 1 18.07 17.49 17.95 controlN/A 34.80 33.72 33.59 (polymer only) NLS0730 1.005 17.76 17.03 17.54NLS0731 0.181 33.67 32.70 32.43

Detection of NLS0109 on Leaves From Plants Grown From Treated SoybeanSeeds.

Soybean seeds were treated with Methylobacterium isolates NLS0109,NS0730, and NLS0731 from 10× frozen glycerol stock to obtain a finalconcentration of 10⁶ CFU/seed. Becker Underwood Flo Rite 1706 polymer isused to improve adhesion. An uninoculated control contained polymer andwater. Seeds were planted in field soil mix, placed in a growth chamberfor approximately two weeks, and watered with unfertilized RO waterevery 1-2 days to keep soil moist. After 2 weeks of growth, true leavesfrom about 9 plants were harvested into sterile tubes. Each treatmenthad at least 2 reps in each experiment, and each experiment was grown atleast 3 times.

DNA from bacteria on the harvested leaves is isolated as follows. Leavesare submerged for 5 minutes in buffer containing 20 mM Tris, 10 mM EDTA,and 0.024% Triton X-100. Tubes are vortexed for 10 minutes, and thensonicated in two 5 minute treatments (10 minutes total). Leaf tissue isremoved, and the remaining liquid centrifuged. The loose pellet is savedand transferred to smaller tubes, while the supernatant is discarded.The sample is centrifuged again, and the final sample obtained as anapproximately 100 ul loose pellet. The 100 ul pellet is used as theinput for DNA extraction using MOBio UltraClean Microbial DNA Extractionkit Cat#12224-250. The average yield of DNA is 50-60 ng/ul in 30ul. Asshown in Table 20, NLS0109 and NLS0730, are detected on leaves harvestedfrom plants grown from soybean seeds treated with the Methylobacteriumstrains using all 3 primer probe sets, as demonstrated by delta Ctvalues of around 5.

TABLE 20 Average of 3 experiments each with 3 biological replicatesSimilarity score to Average Ct Value Treatment NLS0109 Ref1_135566Ref1_135772 Ref1_169470 NLS0109 1.000 35.00 34.67 34.00 control N/A39.67 39.67 39.33 (polymer only) NLS0730 1.005 35.00 35.00 34.00 NLS07310.181 40.00 39.67 40.00

For detection of NLS0109 foliar spray treatment on corn: Untreated cornseeds were planted in field soil in the growth chamber, and watered withnon-fertilized R.O. water. After plants germinated and grew forapproximately 3 weeks, they were transferred to the greenhouse. At V5stage, plants were divided into 3 groups for treatment: foliar spray ofNLS0109, mock foliar spray, and untreated. Plants receiving the foliarspray of NLS0109 were treated with 10× glycerol stock at the rate of71.4 ul per plant using Solo sprayers. This converts to the rate of 10L/acre in the field. Mock treated plants were sprayed with 71.4 ulwater/plant. Untreated plants received no foliar spray treatment. Leaveswere harvested two weeks after foliar spray treatment into sterile tubesand DNA from bacteria on the harvested leaves is isolated as describedabove. Each experiment was grown at least 2 times. As shown in Table 21,NLS0109 is detected on leaves harvested from corn plants treated by afoliar spray application of the Methylobacterium strains using all 3primer probe sets, as demonstrated by delta Ct values of approximately10 between the sample and the negative controls.

TABLE 21 Average Ct Value Treatment Ref1_135566 Ref1_135772 Ref1_169470Control (no 32.43 32.10 31.55 application) Control (mock 35.54 35.3434.80 application) NLS0109 23.36 22.88 22.66 (10 L/acre equivalent)

The above results demonstrate the use of genome specific primers andprobes to detect Methylobacterium strain NLS0109 on various planttissues following treatment with the strains, and provide methods todistinguish NLS0109 from closely related isolates. Similar methods aredeveloped for additional Methylobacterium strains, NLS0020, NLS0017 andNLS0089 using target sequence fragments and primer/probe pairs as shownin Tables 22-27 below.

TABLE 22 Target Fragment Sequences of NLS0020 SEQ ID Fragment NOSequence ref3_25009 21 GCCCTTCTGTCAGGCGATATTGTATAATGGCGTTGCCCCAATAGAAGCAGCCATTCGTGCGAGGGCAGCAGCGACGCTAGGTCGAAAGAGCATCCTAATCTCGATCAAGATGCGACTGAGATTTCTGATGAAAATATCTAGACACAAGCAAAGCTGGTGAAATTACAACGATCATGGCGACAATTGCGGCCAATTCGGCCGGAACTTGAAGGAACATAAAAATGAATATTACAAATATACCGCAAAGCATGTAGAGTTGCTACACCAAGGGTCGGGACGTC CAAAAAAACTCACTGAGGA ref3_2521922 GGAACATAAAAATGAATATTACAAATATACCGCAAAGCATGTAGAGTTGCTACACCAAGGGTCGGGACGTCCAAAAAAACTCACTGAGGAAGTCGACTGGAAGCACGAGGCGCCCCCCCCAGGAGCGGGGCGACCGGCAAGGGGGCCCGCAATTGTCGCCATGATCGACCAGCTTAGGTAGGATCCTCTTTCGACCTAACGAATGGCTGCTTCTATTGGGGCAACGCCATTATACAATATCGCCTGACCATCTGGAACGCGGCCCGGTCCACCGGCAGGTTG GCGACGACAGCGTCGGAG ref1_436122023 CGGCGTCGACCAGCCGGGCGAACTGCTTGGGCATGCTCTCCCGCGACGCCGGCCACAGCCGCGTCCCCGTCCCTCCGCACAGGATCATCGGGTGGATTTGAAAGGCAAAACGGGACATCAGGATAGGCCGCTCAGGCGTTGGCGCTGAGGCGCTTGATGTCGGCGTCGACCATCTCGGTGATCAGCGCCTCGAGGCTGGTCTCGGCCTCCCAGCCGAAGGTCGCCTTGGCCTTGGCGGGGTTGCCCAGCAGCACCTCGACCTCTGCCGGCCGGAACAGCGCCG GGTCGACGATCAGGTGG ref1_460242024 CTGGACATGCGCCCACCCCGGCCAAGTCCGACCGCACCGGCAACCGCTCCTGTAGTCGTCGTCATCGTTCTCACCCCTGAGGCGGAGACCGTCCGCTAACGGGGTGTCTCAAGCAACCGTGGGGCGGAGGAACACGCACGTAGTCGCGTTTCAAGGTTCGCACGAACGCCTCGGCCATGCCGTTGCTCTGCGGGCTCTCCAGCGGCGTCGTTTTTGGCACCAAACCAAGGTCGCGGGCGAAGCGGCGCGTGTCGCGGGGACTGTCAGGAATTTCGTGTGGGG GCGGCCATAGTGGATCCG

TABLE 23 Primer and Probe Sequences for Specific Detection of NLS0020SEQ ID Primer/Probe NO Sequence* NLS0020_ref3_25009_5′ 25CACAAGCAAAGCTGGTGA NLS0020_ref3_25009_3′ 26 AAGATATGCTTTGCGGGTANLS0020_ref3_25009_probe 27 CG A TCATG G C G AC AA TTGNLS0020_ref3_25219_5′ 28 CAATATCGCCTGACAGAAGG NLS0020_ref3_25219_3′ 29CACTTCAACAAAGGCGATCA NLS0020_ref3_25219_probe 30 TTGG C GA C G A C A GCNLS0020_ref1_4361220_5′ 31 ACTGCTTGGGCATGCTCTC NLS0020_ref1_4361220_3′32 CCTATCCTGATGTCCCGTTT NLS0020_ref1_4361220_probe 33 A G GAT C AT C GGG TG G AT T TG NLS0020_ref1_4602420_5′ 34 AGGAACACGCACGTAGTCNLS0020_ref1_4602420_3′ 35 CCACACGAAATTCCTGAC NLS0020_ref1_4602420_probe36 TGG C ACCAA A CCAA *Bold and underlined letters represent theposition of an LNA base

TABLE 24 Target Fragment Sequences of NLS0017 SEQ Fragment ID NOSequence ref4_930 37 GCAAAACGACCTAATAGTTCTACAGCGGCATGCGCCAAGTCAGCGCGGTGAACAGTATACCTGGGAGCAACTTGTCCTCCGAAACCCACATAAAACAAATTACTCCTGGCAGTGCCCAGTCCATCAAAATCGAATACAATATTTCTCGAGGAGGCATCTGTAATAGCCTGCCAAAGCAACAAAGCTATGGCGCCGTTATGACTTTCATTGCTTCTGGTAGACATAAAATAATATGCCGATTTGTGATCCCAAATGTAGAATATTGCCGCATCAATTGCGCCAA GTTTATTTCGGATCGAT ref1_14202138 GGCGCCAACGGTATGATCGCATGATTTTCCTGCGGCATAGCTTGCGGGAATGGCGTATTTGGCGCTCTCCTCAGGAATTTCTAAGGGCATACGCAGGAACTCTACAGCACTTTTACTGGTATTTTGTAGTGACAGCGGAGGAGGCTGGTGCTCAAGGTAATCGTGATGAAGTGATCCGGGCCATTCGGGGCGCGTTTCTAGTCTTTCCAATCCGCGCCCTGTACCACGTATTACGCCGGACCGGTCTGCGCCGCGCCGCCCTCTTGACCGCCCTAAATGTCTAAGA GCGTCTAACAAAGC ref1_142636 39GACGATATCGCTCATCTTCACTGCATTGAAGCTGGTGCCGTACTGCATAGGGATGAAAAAGTGATGCGGATAGACGGCTGACGGGAAAGCGCCTGGTCGATCGAAGACTTTGCTGACGAGGTTGTGGTAGCCCCGGATATAGGCATCGAAGGCCGGGACGTTGATCCCATCCTTTGCCTTATCTTGACTGGCGTCGTCGCGTGCCGTCAGAACGGGCACGTCGCAGGTCATCGAGGCCAGCACCTTGCGGAACACCTGCGTTCCGCCGTTGGGATTATCGACGG CGAACGCGGTGGCCGC

TABLE 25 Primer and Probe Sequences for Specific Detection of NLS0017SEQ Primer/Probe ID NO Sequence* NLS0017_ref4_930_forward 40GTCCTCCGAAACCCACATAAA NLS0017_ref4_930_reverse 41CTACCAGAAGCAATGAAAGTCAT NLS0017_ref4_930_probe 42 TCTGTAATAGCCTGC CAAAGCA NLS0017_ref1_142021_forward 43 GGCTGGTGCTCAAGGTAATNLS0017_ref1_142021_reverse 44 ACATTTAGGGCGGTCAAGAGNLS0017_ref1_142021_probe 45 ATGA A G T GATC C GGGCCATNLS0017_ref1_142636_forward 46 CCGTACTGCATAGGGATGAAANLS0017_ref1_142636_reverse 47 TAAGGCAAAGGATGGGATCAANLS0017_ref1_142636_probe 48 TTGCTGACGAGG TTG TGGTAG *Bold andunderlined letters represent the position of an LNA base

TABLE 26 Target Fragment Sequences of NLS0089 SEQ ID Fragment NOSequence ref1_194299 49 GGAAATCGGCTTCAAGTACGACGTCACGCCGGCCATGCAGGTCACGGGTGCACTGTTCAATCTCGAGCGCGACAACCAGCCGTTCCCCTCGAACGTGGAGTCCGGCCTCGTCCTTGGCGCAGGTCAGACACGCACCCAGGGCGCGGAAATCGGCCTGGCCGGCTATCTAACCGATTGGTGGCAGGTCTTTGGCGGCTACGCTTATACCGAGGCACGCGTACTCTCGCCACTGGAAGACGATGGAGACGTGATCGCAGCAGGTAATCTCGTCGGCAACGTTCC GCTAAATACTTTCAGTCT ref1_19430550 CGGCCTGGCCGGCTATCTAACCGATTGGTGGCAGGTCTTTGGCGGCTACGCTTATACCGAGGCACGCGTACTCTCGCCACTGGAAGACGATGGAGACGTGATCGCAGCAGGTAATCTCGTCGGCAACGTTCCGCTAAATACTTTCAGTCTGTTCAACAAGTTCGATATCAACGAGAATTTCTCCGTTGCTCTGGGCTATTACTATCAGGATGCCAGCTTTGCCTCCTCAGACAATGCAGTGCGTTTGCCAAGTTATTCGCGGTTCGATGGCGGGTTGTTCTATCGA TTCGACGAGTTGAC ref1_194310 51ACGTTCCGCTAAATACTTTCAGTCTGTTCAACAAGTTCGATATCAACGAGAATTTCTCCGTTGCTCTGGGCTATTACTATCAGGATGCCAGCTTTGCCTCCTCAGACAATGCAGTGCGTTTGCCAAGTTATTCGCGGTTCGATGGCGGGTTGTTCTATCGATTCGACGAGTTGACACGCGTTCAGCTTAGCGTCGAGAACATTTTCGACAGGCGTTACATCATCAACTCCAACAACAACAACAACCTCACGCCTGGCGCGCCGAGAACAGTCCGCGTGCAATTGA TCGCTCGGTTCTAAA

TABLE 27 Primer and Probe Sequences for Specific Detection of NLS0089SEQ ID Primer/Probe NO Sequence* NLS0089_ref1_194299_forward 52TTTGGCGGCTACGCTTATAC NLS0089_ref1_194299_reverse 53 AACGTTGCCGACGAGATTACNLS0089_ref1_194299_probe 54 AGACGA T GGAGACGTGATC GC ANLS0089_ref1_194305_forward 55 GGCAACGTTCCGCTAAATACNLS0089_ref1_194305_reverse 56 AAAGCTGGCATCCTGATAGTNLS0089_ref1_194305_probe 57 CGAGA AT TTCTCCGT T GCT C TGNLS0089_ref1_194310_forward 58 CGATGGCGGGTTGTTCTATNLS0089_ref1_194310_reverse 59 AGGCGTGAGGTTGTTGTTNLS0089_ref1_194310_probe 60 AGGCG T TACATCATC A A C TC C A *Bold andunderlined letters represent the position of an LNA base

Example 6 Grey Leaf Spot Field Trials Summer 2017

The effect of Methylobacterium treatment on grey leaf spot disease ofcorn was evaluated in corn field trials. The trials were conducted as arandomized complete block with six replicates. Standard local agronomicpractices for fertilizer application, tillage, row spacing, population,and pest management were used. Standard weed and insect managementpractices were employed and seed was supplied to cooperators pre-treatedwith a standard fungicide/insecticide seed treatment package (Acceleron™2016 Corn, Monsanto, St. Louis, Mo., USA). Foliar fungicide applicationswere only made if specified in the trial protocol and shown in thetables below. When seed was pretreated with Methylobacterium, it wasprovided from a composition comprising from 10⁹-10¹⁰ CFU/ml to providedapproximately 10⁶ CFU/seed. For in furrow or foliar applications, PPFMcompositions were diluted to a concentration of approximately 10⁹ CFU/mland applied at the rates shown in the tables below.

In one trial, treatments with NLS0109 in combination with NLS0017 orNLS0020 were evaluated. Methylobacterium strains were applied either asseed treatment or in furrow applications. Disease severity and yieldresults are provided in Table 28 below. Treatment with NLS0109/NLS0017in furrow provided a 15 bu/acre yield increase over the untreated check.Treatment with NLS0109/NLS0020 as a seed treatment provided a greaterthan 10 bu/acre yield increase over the untreated check. Each of thesetreatments resulted in a yield that was approximately the same (orslightly increased) as for corn treated with Fortix (strobilurin plustriazole).

TABLE 28 Reduction of Grey Leaf Spot Disease with Combinations of PPFMStrains Yield GSF-04 2017 Application (bu/A) GLS Severity UNT 221 6.3NLS 17/109 0.625 liter/A In furrow 236 7.9 NLS 20/109 0.625 liter/A Infurrow 222 7 NLS 17/109 Seed treatment 222 7.9 NLS 20/109 Seed treatment232 7.6 Fortix 4.5 fl oz/A In furrow 228 5.9 Monsoon 4.5 fl oz/A Foliar@ VT 237 6.7 Headline 10 fl. Oz/A Foliar @ VT 254 4.9 Average 231 6.775

In a second trial, NLS0089 and NLS0109 were applied at differentapplication rates alone or in combination with commercial fungicidesalso applied at various rates. All applications were by foliar spray atthe VT stage of development. Disease severity and yield results areprovided in Tables 29 and 30 below.

TABLE 29 NLS 89 and 109 Applied in Combination With Headline(Pyraclostrobin) Application Rate bu/A GLS Severity UNT 241 42.3Headline 0.5 4 oz/A 253 17.7 Headline 1.0 8 oz/A 257 19.4 NLS 89 1.0 2.5Liters/A 232 32.9 Headline 0.5/NLS 89 1.0 4 oz/A 248 20.2 2.5 Liters/AHeadline 1.0/NLS 89 1.0 8 oz/A 249 21.1 2.5 Liters/A NLS 89 0.5 1.25Liters/A 238 41.9 Headline 0.5/NLS 89 0.5 4 oz/A 249 21.4 1.25 Liters/AHeadline 1.0/NLS 89 0.5 8 oz/A 258 18.9 1.25 Liters/A NLS 109 1.0 2.5Liters/A 241 40.6 Headline 0.5/NLS 109 1.0 4 oz/A 254 20.8 2.5 Liters/AHeadline 1.0/NLS 109 1.0 8 oz/A 258 18.4 2.5 Liters/A NLS 109 0.5 1.25Liters/A 229 43.4 Headline 0.5/NLS 109 0.5 4 oz/A 254 23.8 1.25 Liters/AHeadline 1.0/NLS 109 0.5 8 oz/A 243 17.1 1.25 Liters/A

TABLE 30 NLS 89 and 109 Applied in Combination With Monsoon(Tebuconazole) bu/A GLS Severity % UNT 231 49.9 Monsoon 0.5 2 oz/A 23143.0 Monsoon 1.0 4 oz/A 237 43.2 NLS 89 1.0 2.5 Liters/A 231 41.3 Mon0.5/89 1.0 2 oz/A 236 37.8 2.5 Liters/A Mon 1.0/89 1.0 4 oz/A 239 40.82.5 Liters/A NLS 89 0.5 1.25 Liters/A 234 47.6 Mon 0.5/89 0.5 2 oz/A 23245.2 1.25 Liters/A Mon 1.0/89 0.5 4 oz/A 234 34.2 1.25 Liters/A NLS 1091.0 2.5 Liters/A 238 41.1 Mon 0.5/109 1.0 2 oz/A 232 41.3 2.5 Liters/AMon 1.0/109 1.0 4 oz/A 233 40.5 2.5 Liters/A NLS 109 0.5 1.25 Liters/A234 42.7 Mon 0.5/109 0.5 2 oz/A 228 41.5 1.25 Liters/A Mon 1.0/109 0.5 4oz/A 237 42.9 1.25 Liters/A

Example 7 Greenhouse Assay to Determine Activity of MethylobacteriumStrains Against Pythium Species

A greenhouse experiment was conducted is to evaluate the ability ofMethylobacterium strains to suppress Pythium disease in soybean.Methylobacterium treatments were compared to treatments with metalaxylat various rates to determine if Methylobacterium strains can provideprotection beyond and in addition to the level of protection provided bycommonly used agronomic seed treatments. The experiment was designed toallow for growth of plants for in the presence of moderate levels ofPythium. spp. to determine impacts on emergence, % dead seed and rootrot incidence and severity.

Pythium inoculants were prepared by mixing 453 g of parboiled rice and323 mL of distilled water, placing the rice in a vented autoclavableplastic bag, and autoclaving 2×40 minutes with each cycle separated by24 hours. Rice was inoculated with 3-4-day-old Pythium cultures andincubated for 7-10 days at 230C in the dark. The inoculum was then driedfor 2 days.

A 20 ml layer of rice inoculum is placed on soil in cups and additionalsoil layered on top of the inoculum to position the inoculumapproximately 2-3 cm below the seeds to be planted. Water is added tothe soil and 10 seeds are planted in each cup. Domes are placed over thecups and the cups are placed in growth chambers with temperature set at230C, and on a diurnal cycle of 16 h light and 8 h dark. The seeds areincubated for 14 days at 230C. For experiments to assess activity ofMethylobacterium strains against a consortium of 4 Pythium species, theseeds are incubated for 14 days at 130C and for an additional 7 days at230C.

Plants are rated when the first trifoliate is fully emerged and open todetermine total plant weight, number of seedlings and number of deadseeds. Disease incidence is evaluated as percent plants with rottedroots and disease severity as percent rotted root tissue for eachseedling. Results of the analysis are provided in Tables 31-36 below.

Results from inoculation with Pythium sylvaticum at 230C are shown inTables 31-32 below.

TABLE 31 Treatment Emergence % Dead Seed UTC 65 35 0.35 Fl. oz/cwtmetalaxyl 82 18 NLS0089 88 12 NLS0109 87 13

NLS0109 and NLS0089 improved emergence and reduced % dead seed betterthan 7 gm/100 kg metalaxyl active ingredient

TABLE 32 Mean Mean Root Rot Treatment Emergence (%) Severity (%) UTC 6011 1.25 Fl. oz/cwt metalaxyl 78 21 0.75 Fl. oz/cwt metalaxyl 92 18 0.35Fl. oz/cwt metalaxyl 78 17 Bacillus subtilis 83 6 NLS0109 85 5

NLS0109 improved emergence and reduced root rot severity better than 7,15 or 25 gm/100 kg metalaxyl active ingredient

Results from inoculation with Pythium lutarium at 230C are shown inTables 33-34 below.

TABLE 33 Mean Treatment Emergence (%) % Dead Seed UTC 83 17 1.25 Fl.oz/cwt metalaxyl 93 67 0.75 Fl. oz/cwt metalaxyl 97 3 NLS0109 97 3

NLS0109 improved emergence and reduced percent dead seed better than 25gm/100 kg metalaxyl active ingredient and performed as well as 15 gm/100kg metalaxyl active ingredient

TABLE 34 Mean Root Rot Mean Root Rot Treatment Incidence (%) Severity(%) 1.25 Fl. oz/cwt metalaxyl 93 11 0.75 Fl. oz/cwt metalaxyl 95 10 0.35Fl. oz/cwt metalaxyl 95 13 Bacillus subtilis 93 13 NLS0109 92 9

NLS0109 improved emergence and reduced root rot severity better than 7,15 or 25 gm/100 kg metalaxyl active ingredient

Results from inoculation with a consortium of 4 Pythium species atalternating temperature of 130C and 230C are shown in Tables 35-36below. The four species, which are all important pathogens of corn andsoybean, are P. torulosum, P. oopapillum, P. sylvaticum and P. lutarium.

TABLE 35 Mean Mean Dead Treatment Emergence (%) Seed (%) UTC 75 25 0.35Fl. oz/cwt metalaxyl 94 6 NLS0089 100 2 NLS0109 94 2 1.25 Fl. oz/cwtmetalaxyl 94 2 0.75 Fl. oz/cwt metalaxyl 100 2

NLS0109 and NLS0089 improved emergence and NLS0089 reduced % dead seedbetter than 7 gm/100 kg metalaxyl active ingredient.

TABLE 36 Root Rot Root Rot Treatment INC (%) Severity (%) UTC 71 12 1.25Fl. oz/cwt metalaxyl 94 9 0.75 Fl. oz/cwt metalaxyl 100 12 0.35 Fl.oz/cwt metalaxyl 94 13 NLS0089 94 8 NLS0109 90 7

NLS0109 and NLS0089 reduced the incidence and severity of root rot whenapplied to untreated seed better than 7, 15 or 25 gm/100 kg metalaxylactive ingredient

Example 8 Analysis of Effect of Methylobacterium Strains on Rhizoctoniaand Fusarium Soil Diseases

The effect of Methylobacterium treatment on emergence was evaluated insoy and corn field trials. Improved emergence is evidence of activity ofthe Methylobacterium treatment against common Fusarium and Rhizoctoniasoil pathogens. The trials were conducted as a randomized complete blockwith four replicates. Standard local agronomic practices for fertilizerapplication, tillage, row spacing, population, and pest management wereused. Standard weed and insect management practices were employed andseed was supplied to cooperators pre-treated with a standardfungicide/insecticide seed treatment. Seed treatment compositions forsoybean contained ipconazole, metalaxyl, imidacloprid and a seed coatingin addition to the Methylobacterium strains. Seed treatment compositionsfor corn contained ipconazole, metalaxyl, trifloxystrobin andclothianidin. Stand counts were made at 21 days after first emergence orabout 27 days after planting. Effect of Methylobacterium treatment onemergence in corn was evaluated with Methylobacterium provided as a seedtreatment or by in furrow application. Results are shown in Tables 37-39below.

TABLE 37 Soybean Emergence Treatment Stand Count NLS 20/109 118432 NLS109 118170 Integral ™ (B. subtilis) 114456 PPST 2030 114431 NLS 64/109112965 UNT 110376 NLS 17/109 109652 NLS 20/109 105465

Seed treatment with NLS0109 improved soybean stand count in comparisonto treatment with Integral™ (B. subtilis, BASF), PPST 2030 (PioneerPremium Seed Treatment) and untreated control.

TABLE 38 Corn Emergence following Methylobacterium seed treatmentTreatment Stand Count NLS 17/109 34272 UNT 34141 NLS 20/109 32638

TABLE 39 Corn Emergence following Methylobacterium in furrow applicationTreatment Stand Count NLS 17/109 34445 NLS 20/109 33302 UNT 33186

Seed treatment with NLS0109 in combination with NLS0017 improved cornemergence in comparison to untreated control.

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The inclusion of various references herein is not to be construed as anyadmission by the Applicant that the references constitute prior art.Applicants expressly reserve their right to challenge any allegations ofunpatentability of inventions disclosed herein over the referencesincluded herein.

Having illustrated and described the principles of the presentdisclosure, it should be apparent to persons skilled in the art that thedisclosure can be modified in arrangement and detail without departingfrom such principles.

Although the materials and methods of this disclosure have beendescribed in terms of various embodiments and illustrative examples, itwill be apparent to those of skill in the art that variations can beapplied to the materials and methods described herein without departingfrom the concept, spirit and scope of the disclosure. All such similarsubstitutes and modifications apparent to those skilled in the art aredeemed to be within the spirit, scope and concept of the disclosure asdefined by the appended claims or otherwise disclosed herein.

1.-90. (canceled)
 91. A method to detect the presence ofMethylobacterium strain NL0109, NLS0020, NLS0017, or NLS0089 in asample, wherein said method comprises detecting the presence in thesample of a nucleic acid comprising or located within SEQ ID NO:9-11,21-24, 37-39 or 49-51.
 92. The method of claim 91, wherein the detectingof the sequence comprises a polymerase chain reaction, branched DNA,ligase chain reaction, transcription mediated amplification (TMA),nucleic acid sequence-based amplification (NASBA), nanopore-massspectroscopy, hybridization, or direct sequencing based method, or anycombination thereof.
 93. The method of claim 91, said detectioncomprises the steps of (i) contacting the sample with a DNA primer pair,wherein said primer pair comprises forward and reverse primers foramplification of a DNA fragment comprising or located within SEQ IDNO:9-11, 21-24, 37-39 or 49-51, thereby generating a DNA fragment, (ii)contacting said DNA fragment with a probe specific for the presence ofsaid DNA fragment, and (iii) comparing the results of said contactingwith positive and negative controls to determine the presence of SEQ IDNO:9-11, 21-24, 37-39 or 49-51 in said sample.
 94. The method of claim91 wherein said sample is a plant material that was treated with one ormore of Methylobacterium strains selected from NL0109, NLS0020, NLS0017,or NLS0089.
 95. The method of claim 93 wherein said plant material isleaves, roots or seeds.
 96. The method of claim 93 wherein the plantmaterial is a processed plant product from a plant treated with one ormore Methylobacterium strains selected from NLS0109, NLS0020, NLS0017,or NLS0089.
 97. The method of claim 91 wherein said sample is a soilsample.
 98. A method of treating a plant or plant part, wherein saidmethod comprises spraying, coating, partially coating, immersing, and/orimbibing the plant or plant part with a composition comprising afermentation product comprising a Methylobacterium strain selected fromthe group consisting of NLS0109 (NRRL B-67340) and a derivative ofNLS0109 having a sequence of any one of SEQ ID NOS: 9-11.
 99. The methodof claim 98 wherein said Methylobacterium is at a concentration of atleast 5×10⁷ colony-forming units per gram or 5×10⁷ colony-forming unitsper milliliter.
 100. The method of claim 98 wherein said compositionfurther comprises a carrier.
 101. The method of claim 98 wherein saidcomposition further comprises at least one of: (i) an agriculturallyacceptable excipient comprising at least one component selected from thegroup consisting of woodflours, clays, activated carbon, diatomaceousearth, fine-grain inorganic solids, calcium carbonate, calciumbentonite, kaolin, china clay, talc, perlite, mica, vermiculite,silicas, quartz powder, montmorillonite, and mixtures thereof, and (ii)an agriculturally acceptable adjuvant comprising at least one componentselected from the group consisting polyvinyl acetates, polyvinyl acetatecopolymers, hydrolyzed polyvinyl acetates, polyvinylpyrrolidone-vinylacetate copolymer, polyvinyl alcohols, polyvinyl alcohol copolymers,polyvinyl methyl ether, polyvinyl methyl ether-maleic anhydridecopolymer, waxes, latex polymers, celluloses, ethylcelluloses,methylcelluloses, hydroxy methylcelluloses, hydroxypropylcellulose,hydroxymethylpropylcelluloses, polyvinyl pyrrolidones, alginates,dextrins, malto-dextrins, polysaccharides, fats, oils, proteins, karayagum, jaguar gum, tragacanth gum, polysaccharide gums, mucilage, gumarabics, shellacs, vinylidene chloride polymers and copolymers,soybean-based protein polymers and copolymers, lignosulfonates, acryliccopolymers, starches, polyvinylacrylates, zeins, gelatin,carboxymethylcellulose, chitosan, polyethylene oxide, acrylamidepolymers and copolymers, polyhydroxyethyl acrylate, methylacrylamidemonomers, alginate, polychloroprene, and syrups or mixtures thereof.102. The method of claim 101 wherein the composition comprises anagriculturally acceptable excipient selected from the group consistingof woodflours, clays, activated carbon, diatomaceous earth, fine-graininorganic solids, calcium carbonate, calcium bentonite, kaolin, chinaclay, talc, perlite, mica, vermiculite, silicas, quartz powder,montmorillonite, and mixtures thereof.
 103. The method of claim 98 theplant part coated with the Methylobacterium composition is selected fromthe group consisting of a leaf, a stem, a flower, a fruit, a coleoptile,a root, a tuber, and a seed.
 104. The method of claim 98 wherein saidplant is selected from the group consisting of corn, a Brassica sp.,alfalfa, rice, rye, sorghum, millet, sunflower, safflower, soybean,tobacco, potato, peanuts, cotton, sweet potato, cassava, coffee,coconut, pineapple, citrus trees, cocoa, tea, banana, avocado, fig,guava, mango, olive, papaya, cashew, macadamia, almond, sugar beets,sugarcane, oats, barley, tomatoes, lettuce, green beans, lima beans,peas, cucurbits, ornamentals, and conifers.
 105. The method of claim 98wherein said plant is a cereal plant selected from the group consistingof rice, wheat, corn, barley, millet, sorghum, oat, and rye.
 106. Themethod of claim 98 wherein said plant is a turfgrass plant selected fromthe group consisting of annual bluegrass, annual ryegrass, Canadabluegrass, fescue, bentgrass, wheatgrass, Kentucky bluegrass, orchardgrass, ryegrass, redtop, Bermuda grass, St. Augustine grass, and zoysiagrass.
 107. The method of claim 98 wherein said composition furthercomprises a beneficial microorganism other than said Methylobacteriumstrain.
 108. The method of claim 107 wherein said beneficialmicroorganism is selected from the group consisting of a Bacillus sp., aPseudomonas sp., a Coniothyrium sp., a Pantoea sp., a Streptomyces sp.,a Trichoderma sp., a Rhizobium sp. and a Bradyrhizobium sp.
 109. Themethod of claim 98 wherein the composition further comprises a secondMethylobacterium strain selected from the group consisting of NLS0017(NRRL B-50931), NLS0020 (NRRL B-50930), and a Methylobacterium strainderived from or related to NLS0017 or NLS0020 and having a sequence ofany one of SEQ ID NOS: 21-24 or 37-39.
 110. The method of claim 98wherein said Methylobacterium strain is NLS0109 (NRRL B-67340).
 111. Themethod of claim 109 wherein said second Methylobacterium strain isNLS0017 (NRRL B-50931) or NLS0020 (NRRL B-50930).
 112. The method ofclaim 98 wherein said plant is a cucurbit plant selected from the groupconsisting of cucumber, cantaloupe, and musk melon.